RAGE/AGER Antibody (175410)

Detection of RAGE by Immunohistochemistry

Double-positive cells in Sftpc-CreER; LSL-KrasG12D-induced lung adenocarcinoma are tumor-propagating cells and forms tumors upon transplantation. A, Experimental outline for growing cells in 3D organoid culture. B, FACS scatter plot showing single (type I or type II) and double-positive population sorted from Sftpc-CreER; LSL-KrasG12D; TdTomato mice. Type I (TdTm−Pdpn+), type II (TdTm+ Pdpn−), and double-positive (type I/II+; TdTm+Pdpn+) fractions were sorted. C, Brightfield images of 3D organoid cultures from different indicated sorted populations on day 14. Colony number (D) and colony size (E) was quantified using ImageJ software. F, 3D organoid culture from double-positive cells on day 14 in red channel. G, Immunofluorescent image on day 14 of organoid culture showing double-positive cells differentiate into type I and type II cells. Ki67 (H) and TTF1 (I) staining of 3D organoid culture. J, Representative H&E staining of tumors sections which were obtained after subcutaneous transplantation of double-positive (type I/II+) cells. Right panel shows higher magnification of boxed area in left panels. K, Representative IHC images of tumor sections for TTF1. Right panel shows higher magnification of boxed area in left panels. L, Representative immunofluorescent image for in red channel showing Tdtomato expression in tumor obtained after transplantation. M, Representative IF images for Sftpc and Rage staining. Right panel shows higher magnification of boxed area in left panels. Results shown in B, F, G, H, I, J, K, L, and M are representative of three independent experiments. D and E represent data from three independent experiments. Error bars represent the mean ± SEM values and significance is defined as *, P ≤ 0.05; **, P ≤ 0.01; and ***, P ≤ 0.001. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37882674), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of RAGE by Immunohistochemistry

SARS-CoV-2 distribution and viral target cells in the lungs from each mouse model. (A) SARS-CoV-2 distribution in the lungs analysed using in situ hybridisation at 2, 5, and 7 dpi with 1 × 105 PFU SARS-CoV-2. (B) Representative confocal immunofluorescence micrographs of the lungs of K18-hACE2 (left), SFTPB-hACE2 (middle), and SCGB1A1-hACE2 (right) mice from the PBS-treated control group (top) and SARS-CoV-2-infected group (bottom) at 2 dpi. SARS-CoV-2 N-protein-positive cells are stained red, and type 1 alveolar (Ager), type 2 alveolar (Lamp3), or club (SCGB1A1) cells are stained green. Nuclei of cells are stained blue with DAPI. The scale bars are 100 μm. (C) Quantification of the percentage of infected cells from the lungs of K18-hACE2 (left), SFTPB-hACE2 (middle), and SCGB1A1-hACE2 (right) mice analysed from the confocal immunofluorescence micrographs shown in (B). Lung sections from SARS-CoV-2-infected mice at 2 dpi stained with anti-SARS-CoV-2 N-protein antibody and one of anti-Ager, anti-Lamp3, anti-SCGB1A1, anti-Muc5ac, or anti-tubulin antibodies. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36457995), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of RAGE by Immunohistochemistry

Sustained activation of YAP/TAZ drives the fate conversion of secretory cells into AT1 cells via DATP cell statesAExperimental design for isolating Scgb1a1 lineage‐labeled cells from control and Lats1/2 dKO lungs at indicated time points post tamoxifen treatment. Isolated cells were subjected to scRNA‐seq analysis.BClusters of Scgb1a1 lineage‐labeled cells from 10XGenomics 3’ single‐cell RNA sequencing (scRNA‐seq) analysis visualized by Uniform Manifold Approximation and Projection (UMAP), assigned by specific colors at day 5 and 14 post tamoxifen treatment. The number of cells in the individual cluster is depicted in the figure.CDistribution of each cluster across indicated time points after tamoxifen treatment.DBar graph showing the percentage of secretory, DATPs, and AT1 cell clusters in control and Lats1/2 dKO lungs at day 5 and 14 post tamoxifen treatment.EBlob plot depicting selected marker genes in cell clusters. Dot size encodes the percentage of cells expressing the gene, and color encodes the average per cell gene expression level.FPseudotime ordering of Scgb1a1 lineage‐labeled cells colored by cluster assignment (right) according to diffusion pseudotime (DPT, left) order.G, HRepresentative IF images of control and Lats1/2 dKO airways at indicated time points post tamoxifen treatment. Tomato (for Scgb1a1 lineage, red), CLDN4 (for DATPs, white), CC10 (G, green), AGER (H, for AT1 cell, green), and DAPI (blue). Scale bar, 100 μm.IQuantification of Scgb1a1 lineage‐labeled tdTomato+CLDN4+ DATPs in (G and H). Data are presented as mean ± SEM (n = 5 mice for each genotype). ***P < 0.001 (Student’s t‐test). Image collected and cropped by CiteAb from the following open publication (https://link.springer.com/10.15252/embj.2021109365), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of RAGE by Western Blot

L-4F treatment decreases RAGE/ED1/HMGB-1, and PAI-1 in the ischemic brain in T2DM mice 4 days after MCAo. (A–D) HMGB-1 immunostaining and quantitative data; (E,F) RAGE/ED1/HMGB-1 WB assay and quantitative data; (G,H) TNF alpha /PAI-1 WB assay and quantitative data; (I) TNF alpha /PAI-1 RT-PCR data. Scale bar in (A–C) = 50 μm; ∗p < 0.05, n = 11/group in (A–D); n = 6/group in (A–I). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/31708728), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of RAGE by Immunocytochemistry/ Immunofluorescence

Immunofluorescent staining revealed increased RAGE expression in TG mouse lungs ((F,K) vs. A) and diminished localization of NKX2.1 ((G,L) vs. B) and alpha smooth muscle actin (ACTA 2; (H,M) vs. C). DAPI counterstained images (D,I,N) and merged images that contains RAGE, NKX2.1, ACTA 2, and DAPI (E,J,O) are also depicted. All images are at 400× original magnification and scale bars represent 50 μm. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37489334), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of RAGE by Western Blot

(A) Doxycycline (Dox)-inducible expression of RAGE in transgenic (TG) mice required a reverse tetracycline transactivator (rtTA) localized to respiratory epithelial cells via the human surfactant protein (hSP)-C promoter. The rtTA complexed with Dox and activated Tet-inducible RAGE expression. (B) Representative immunoblotting revealed increased expression of RAGE in RAGE TG mice fed Dox compared to Dox-exposed control animals. Actin was used as a loading control. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37489334), licensed under a CC-BY license. Not internally tested by R&D Systems.