Human EGLN2/PHD1 Antibody

Detection of Human EGLN2/PHD1 by Western Blot.

Western blot shows lysates of MDA-MB-231 human breast cancer cell line. PVDF Membrane was probed with 1 µg/mL of Human EGLN2/PHD1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6394) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for EGLN2/PHD1 at approximately 48 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.

Detection of Mouse EGLN2/PHD1 by Immunohistochemistry

NQO1 promotesin vivo tumour growth.(a) RKO/pNQO1, RKO/shCont and RKO/pshNQO1 cells were injected subcutaneously into the right flank of athymic, 7-week-old female BALB/C nude mice, and tumour growth was assessed. Tumour volume (TV) was calculated by using the following formula: TV=length × (width)2 × 0.5. Each group contained 12 animals. (**P<0.01 with unpaired t-test). (b) Immunohistochemical analyses of RKO/pNQO1, RKO/shCont, and RKO/pshNQO1 xenograft tumours. The sections were stained for NQO1, HIF-1 alpha, proliferation (Ki67) and apoptosis (CC3) using 3,3′-DAB. Scale bar, 50 μm. (c) Quantification of NQO1, proliferative marker Ki67 and apoptotic marker CC3 in RKO/pNQO1, RKO/shCont and RKO/pshNQO1 xenograft tumours (n=3 each group). n=5 in each tumour. Two-tail t-test. **P<0.01, *P<0.05. #, not significant. All error bars represent the mean±s.e.m. (d) RKO/pshCont1/pshCont2, RKO/pNQO1/pshCont2 and RKO/pNQO1/pshHIF-1 alpha injected subcutaneously into the right flank of athymic, 7-week-old female BALB/C nude mice, and tumour growth was assessed. Tumour volume (TV) was calculated by using the following formula: TV=length × (width)2 × 0.5. Each group contained 10 animals (*P<0.05 with unpaired t-test). (e) Immunohistochemical analyses of RKO/pshCont1/pshCont2, RKO/pNQO1/pshCont2 and RKO/pNQO1/pshHIF-1 alpha xenograft tumours for HIF-1 alpha, proliferation (Ki67), apoptosis (CC3) and vasculature (CD34) using 3,3′-DAB. Arrowheads denote blood vessels. Scale bar, 50 μm. (f) RKO/pNQO1/pshCont or RKO/pNQO1/pshHIF-1 alpha xenograft tumours (n=3–5 each group) were quantified for proliferation (Ki67), apoptosis (CC3) and vascularization (CD34). n=5 in each tumour. Two tailed t-test. **P<0.01, *P<0.05. #, not significant. All error bars represent the mean±s.e.m. (g) Schematic model showing how NQO1 stabilizes HIF-1 alpha in cancer cells. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/27966538), licensed under a CC-BY license. Not internally tested by

Detection of Mouse EGLN2/PHD1 by Immunohistochemistry

NQO1 promotesin vivo tumour growth.(a) RKO/pNQO1, RKO/shCont and RKO/pshNQO1 cells were injected subcutaneously into the right flank of athymic, 7-week-old female BALB/C nude mice, and tumour growth was assessed. Tumour volume (TV) was calculated by using the following formula: TV=length × (width)2 × 0.5. Each group contained 12 animals. (**P<0.01 with unpaired t-test). (b) Immunohistochemical analyses of RKO/pNQO1, RKO/shCont, and RKO/pshNQO1 xenograft tumours. The sections were stained for NQO1, HIF-1 alpha, proliferation (Ki67) and apoptosis (CC3) using 3,3′-DAB. Scale bar, 50 μm. (c) Quantification of NQO1, proliferative marker Ki67 and apoptotic marker CC3 in RKO/pNQO1, RKO/shCont and RKO/pshNQO1 xenograft tumours (n=3 each group). n=5 in each tumour. Two-tail t-test. **P<0.01, *P<0.05. #, not significant. All error bars represent the mean±s.e.m. (d) RKO/pshCont1/pshCont2, RKO/pNQO1/pshCont2 and RKO/pNQO1/pshHIF-1 alpha injected subcutaneously into the right flank of athymic, 7-week-old female BALB/C nude mice, and tumour growth was assessed. Tumour volume (TV) was calculated by using the following formula: TV=length × (width)2 × 0.5. Each group contained 10 animals (*P<0.05 with unpaired t-test). (e) Immunohistochemical analyses of RKO/pshCont1/pshCont2, RKO/pNQO1/pshCont2 and RKO/pNQO1/pshHIF-1 alpha xenograft tumours for HIF-1 alpha, proliferation (Ki67), apoptosis (CC3) and vasculature (CD34) using 3,3′-DAB. Arrowheads denote blood vessels. Scale bar, 50 μm. (f) RKO/pNQO1/pshCont or RKO/pNQO1/pshHIF-1 alpha xenograft tumours (n=3–5 each group) were quantified for proliferation (Ki67), apoptosis (CC3) and vascularization (CD34). n=5 in each tumour. Two tailed t-test. **P<0.01, *P<0.05. #, not significant. All error bars represent the mean±s.e.m. (g) Schematic model showing how NQO1 stabilizes HIF-1 alpha in cancer cells. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/27966538), licensed under a CC-BY license. Not internally tested by