Human IFITM1 Antibody

Detection of Human IFITM1 by Western Blot. Western blot shows lysates of Jurkat human acute T cell leukemia cell line, TF-1 human erythroleukemic cell line, human placenta tissue. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human IFITM1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4827) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF019). A specific band was detected for IFITM1 at approximately 17 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.

IFITM1 in K562 Human Cell Line. IFITM1 was detected in immersion fixed K562 human chronic myelogenous leukemia cell line using Goat Anti-Human IFITM1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4827) at 1.7 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to plasma membranes and cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Detection of IFITM1 by Western Blot IFITM1/2/3 triple knockout enhances KSHV and RRV infection in A549 cells and HFF. A549 cells (A), HFF (B), and HUVEC (C) were transduced with lentiviral vectors encoding Cas9 and the sgRNAs shown in Fig. 2. (A to C, left panels) IFITM knockout (sgIFITM1/2/3-a, sgIFITM1/2/3-b) or control (sgNT-a, sgNT-b) cells treated with IFN-alpha (5,000 U/ml) or H2O (control) and infected with KSHV-GFP, RRV-YFP, IAV lentiviral pseudotype (IAV-LP), or MLV lentiviral pseudotype (MLV-LP). Infection was measured using flow cytometry to detect expression of the fluorescent reporter gene. The graph shows individual data points representing averaged values for GFP+/YFP+ cells of either two nontargeting (sgNT-a, sgNT-b) or IFITM1/2/3 knockout (sgIFITM1/2/3-a, sgIFITM1/2/3-b) transduced cells and floating bars representing the mean averaged from results of four independent experiments for A549 cells and HFF (A and B) and three independent experiments for HUVEC (C). Infections for each single experiment were performed in triplicate for each condition. Data points from the same experiment are labeled with identical symbols. The different sgRNAs were treated as biological replicates within each experiment. Statistical significance was determined by two‐way analysis of variance (ANOVA), and P values were corrected for all possible multiple comparisons within one family by Tukey’s method (nonsignificant [ns], P > 0.05; *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001; ****, P ≤ 0.0001). (A to C, right panels) Representative Western blots of IFITM knockout (sgIFITM1/2/3-a or sgIFITM1/2/3-b) or control (sgNT-a or sgNT-b) cells treated with IFN-alpha (5,000 U/ml) or H2O. Indicated IFITM expression was detected with antibodies shown in Fig. 1A; GAPDH served as a loading control. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/34933450), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of IFITM1 by Western Blot KSHV induces IFITM1, IFITM2, and IFITM3 expression in A549 cells. (A) Western blot of 293T cells transduced with pQCXIP constructs to express IFITM1 to -3 or pQCXIP (empty vector). IFITMs were detected using the respective IFITM antibody, and GAPDH served as a loading control. (B and C) Fluorescence microscopy images (scale bar, 200 μm) (B) and Western blot analysis (C) of A549 cells infected with KSHV-GFP or RRV-YFP or treated with H2O or IFN-alpha (5,000 U/ml) for the indicated time and harvested using SDS sample buffer. IFITM expression was detected with antibodies shown in panel A. MxA served as control for IFN-stimulated gene induction; GAPDH served as a loading control. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/34933450), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of IFITM1 by Western Blot Overexpression of IFITM1 inhibits KSHV and RRV infection in a cell-specific manner. A549 cells (A), HFF (B), 293T cells (C), and SLK cells (D) were transduced with pQCXIP constructs to express IFITM1-3 or pQCXIP (empty vector). (A to D, left panels) IFITM-overexpressing cells were infected with KSHV-GFP, RRV-YFP, IAV lentiviral pseudotype (IAV-LP), or MLV lentiviral pseudotype (MLV-LP). Infection was measured using flow cytometry to detect expression of the fluorescent reporter genes. The data show values normalized to pQCXIP empty vector, which was set to 100%, and the error bars represent the standard error of the mean of results of four independent experiments, each performed in triplicate. Statistical significance was determined by ordinary two-way ANOVA, and P values were corrected for multiple comparisons by Dunnett’s method (ns, P > 0.05; *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001; ****, P ≤ 0.0001). (A to C, right panels) Representative Western blots of IFITM-overexpressing cells. Expression of myc-tagged IFITMs was determined using anti-myc antibody; GAPDH served as a loading control. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/34933450), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of IFITM1 by Western Blot Overexpression of IFITM1 inhibits KSHV and RRV infection in a cell-specific manner. A549 cells (A), HFF (B), 293T cells (C), and SLK cells (D) were transduced with pQCXIP constructs to express IFITM1-3 or pQCXIP (empty vector). (A to D, left panels) IFITM-overexpressing cells were infected with KSHV-GFP, RRV-YFP, IAV lentiviral pseudotype (IAV-LP), or MLV lentiviral pseudotype (MLV-LP). Infection was measured using flow cytometry to detect expression of the fluorescent reporter genes. The data show values normalized to pQCXIP empty vector, which was set to 100%, and the error bars represent the standard error of the mean of results of four independent experiments, each performed in triplicate. Statistical significance was determined by ordinary two-way ANOVA, and P values were corrected for multiple comparisons by Dunnett’s method (ns, P > 0.05; *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001; ****, P ≤ 0.0001). (A to C, right panels) Representative Western blots of IFITM-overexpressing cells. Expression of myc-tagged IFITMs was determined using anti-myc antibody; GAPDH served as a loading control. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/34933450), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of IFITM1 by Western Blot IFITM1/2/3 triple knockout enhances KSHV and RRV infection in A549 cells and HFF. A549 cells (A), HFF (B), and HUVEC (C) were transduced with lentiviral vectors encoding Cas9 and the sgRNAs shown in Fig. 2. (A to C, left panels) IFITM knockout (sgIFITM1/2/3-a, sgIFITM1/2/3-b) or control (sgNT-a, sgNT-b) cells treated with IFN-alpha (5,000 U/ml) or H2O (control) and infected with KSHV-GFP, RRV-YFP, IAV lentiviral pseudotype (IAV-LP), or MLV lentiviral pseudotype (MLV-LP). Infection was measured using flow cytometry to detect expression of the fluorescent reporter gene. The graph shows individual data points representing averaged values for GFP+/YFP+ cells of either two nontargeting (sgNT-a, sgNT-b) or IFITM1/2/3 knockout (sgIFITM1/2/3-a, sgIFITM1/2/3-b) transduced cells and floating bars representing the mean averaged from results of four independent experiments for A549 cells and HFF (A and B) and three independent experiments for HUVEC (C). Infections for each single experiment were performed in triplicate for each condition. Data points from the same experiment are labeled with identical symbols. The different sgRNAs were treated as biological replicates within each experiment. Statistical significance was determined by two‐way analysis of variance (ANOVA), and P values were corrected for all possible multiple comparisons within one family by Tukey’s method (nonsignificant [ns], P > 0.05; *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001; ****, P ≤ 0.0001). (A to C, right panels) Representative Western blots of IFITM knockout (sgIFITM1/2/3-a or sgIFITM1/2/3-b) or control (sgNT-a or sgNT-b) cells treated with IFN-alpha (5,000 U/ml) or H2O. Indicated IFITM expression was detected with antibodies shown in Fig. 1A; GAPDH served as a loading control. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/34933450), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of IFITM1 by Western Blot IFITM1/2/3 triple knockout enhances KSHV and RRV infection in A549 cells and HFF. A549 cells (A), HFF (B), and HUVEC (C) were transduced with lentiviral vectors encoding Cas9 and the sgRNAs shown in Fig. 2. (A to C, left panels) IFITM knockout (sgIFITM1/2/3-a, sgIFITM1/2/3-b) or control (sgNT-a, sgNT-b) cells treated with IFN-alpha (5,000 U/ml) or H2O (control) and infected with KSHV-GFP, RRV-YFP, IAV lentiviral pseudotype (IAV-LP), or MLV lentiviral pseudotype (MLV-LP). Infection was measured using flow cytometry to detect expression of the fluorescent reporter gene. The graph shows individual data points representing averaged values for GFP+/YFP+ cells of either two nontargeting (sgNT-a, sgNT-b) or IFITM1/2/3 knockout (sgIFITM1/2/3-a, sgIFITM1/2/3-b) transduced cells and floating bars representing the mean averaged from results of four independent experiments for A549 cells and HFF (A and B) and three independent experiments for HUVEC (C). Infections for each single experiment were performed in triplicate for each condition. Data points from the same experiment are labeled with identical symbols. The different sgRNAs were treated as biological replicates within each experiment. Statistical significance was determined by two‐way analysis of variance (ANOVA), and P values were corrected for all possible multiple comparisons within one family by Tukey’s method (nonsignificant [ns], P > 0.05; *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001; ****, P ≤ 0.0001). (A to C, right panels) Representative Western blots of IFITM knockout (sgIFITM1/2/3-a or sgIFITM1/2/3-b) or control (sgNT-a or sgNT-b) cells treated with IFN-alpha (5,000 U/ml) or H2O. Indicated IFITM expression was detected with antibodies shown in Fig. 1A; GAPDH served as a loading control. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/34933450), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of IFITM1 by Western Blot KSHV induces IFITM1, IFITM2, and IFITM3 expression in A549 cells. (A) Western blot of 293T cells transduced with pQCXIP constructs to express IFITM1 to -3 or pQCXIP (empty vector). IFITMs were detected using the respective IFITM antibody, and GAPDH served as a loading control. (B and C) Fluorescence microscopy images (scale bar, 200 μm) (B) and Western blot analysis (C) of A549 cells infected with KSHV-GFP or RRV-YFP or treated with H2O or IFN-alpha (5,000 U/ml) for the indicated time and harvested using SDS sample buffer. IFITM expression was detected with antibodies shown in panel A. MxA served as control for IFN-stimulated gene induction; GAPDH served as a loading control. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/34933450), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of IFITM1 by Western Blot Overexpression of IFITM1 inhibits KSHV and RRV infection in a cell-specific manner. A549 cells (A), HFF (B), 293T cells (C), and SLK cells (D) were transduced with pQCXIP constructs to express IFITM1-3 or pQCXIP (empty vector). (A to D, left panels) IFITM-overexpressing cells were infected with KSHV-GFP, RRV-YFP, IAV lentiviral pseudotype (IAV-LP), or MLV lentiviral pseudotype (MLV-LP). Infection was measured using flow cytometry to detect expression of the fluorescent reporter genes. The data show values normalized to pQCXIP empty vector, which was set to 100%, and the error bars represent the standard error of the mean of results of four independent experiments, each performed in triplicate. Statistical significance was determined by ordinary two-way ANOVA, and P values were corrected for multiple comparisons by Dunnett’s method (ns, P > 0.05; *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001; ****, P ≤ 0.0001). (A to C, right panels) Representative Western blots of IFITM-overexpressing cells. Expression of myc-tagged IFITMs was determined using anti-myc antibody; GAPDH served as a loading control. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/34933450), licensed under a CC-BY license. Not internally tested by R&D Systems.