Human FoxF1 Antibody

Detection of Human FoxF1 by Western Blot. Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line, Jurkat human acute T cell leukemia cell line, and PC-3 human prostate cancer cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human FoxF1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4798) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for FoxF1 at approximately 50 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of Human FoxF1 by Western Blot. Western blot shows lysates of WI-38 human lung fibroblast cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human FoxF1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4798) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for FoxF1 at approximately 44 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of Human FoxF1 by Western Blot Loss of FOXF1 promotes fundamental cellular processes in LR-MSCs. (A) mRNA was isolated from fibrotic and non-fibrotic LR-MSCs derived from bronchoalveolar lavage fluid of transplant patients, and FOXF1 expression was analyzed by real-time PCR. Values: Means ± SEM; n = 9 (non-Fib-MSCs); n = 8 (Fib-MSCs); **p < 0.0034. (B) Protein lysates from fibrotic and non-fibrotic LR-MSCs were analyzed for FOXF1 and GAPDH by western blotting. Graph shows densitometry analyses of these immunoblots. Values: Means ± SEM; n = 16; **p < 0.0086. (C) LR-MSCs were transfected with scrambled or FOXF1-specific siRNA and confirmed by real-time PCR. Values: Means ± SEM; n = 7; ***p < 0.0005. (D) Protein lysates from (A) were subjected to immunoblotting with antibodies against FOXF1 and GAPDH. (E–I) Gene regulation due to FOXF1 silencing was analyzed by Affymetrix gene array in 3 lines of LR-MSCs. Data reflects fold changes ≥ 1.5, and an adjusted p < 0.01. (E) Diagram showing the number of upregulated and downregulated genes. (F) Gene–gene interaction network (using STRING database) showing associations due to FOXF1-silencing. (G–I) Heatmaps showing two-fold Log changes are presented for positive regulation of cell cycle ((G) GO:0045787), inflammatory response ((H) GO:0006954), and regulation of cell migration ((I) GO:0030334). Note: Full length blots for Fig. 1B and Fig. 1D are provided in Supplementary Fig. S1 and S2. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/33277571), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human FoxF1 by Western Blot Loss of FOXF1 promotes fundamental cellular processes in LR-MSCs. (A) mRNA was isolated from fibrotic and non-fibrotic LR-MSCs derived from bronchoalveolar lavage fluid of transplant patients, and FOXF1 expression was analyzed by real-time PCR. Values: Means ± SEM; n = 9 (non-Fib-MSCs); n = 8 (Fib-MSCs); **p < 0.0034. (B) Protein lysates from fibrotic and non-fibrotic LR-MSCs were analyzed for FOXF1 and GAPDH by western blotting. Graph shows densitometry analyses of these immunoblots. Values: Means ± SEM; n = 16; **p < 0.0086. (C) LR-MSCs were transfected with scrambled or FOXF1-specific siRNA and confirmed by real-time PCR. Values: Means ± SEM; n = 7; ***p < 0.0005. (D) Protein lysates from (A) were subjected to immunoblotting with antibodies against FOXF1 and GAPDH. (E–I) Gene regulation due to FOXF1 silencing was analyzed by Affymetrix gene array in 3 lines of LR-MSCs. Data reflects fold changes ≥ 1.5, and an adjusted p < 0.01. (E) Diagram showing the number of upregulated and downregulated genes. (F) Gene–gene interaction network (using STRING database) showing associations due to FOXF1-silencing. (G–I) Heatmaps showing two-fold Log changes are presented for positive regulation of cell cycle ((G) GO:0045787), inflammatory response ((H) GO:0006954), and regulation of cell migration ((I) GO:0030334). Note: Full length blots for Fig. 1B and Fig. 1D are provided in Supplementary Fig. S1 and S2. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/33277571), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human FoxF1 by Western Blot ATX-dependent cell migration in FOXF1-silenced LR-MSCs. (A) LR-MSCs were transfected with scrambled or FOXF1-specific siRNA. 24 h later, these LR-MSCs were transfected with scrambled or ATX-specific siRNA. Immunoblotting was performed to confirm RNAi-mediated FOXF1 and ATX silencing efficacy. n = 5 per group. (B) Migration assay was conducted in LR-MSCs transfected with scrambled or siRNA specific to FOXF1, ATX, or both FOXF1- and ATX-specific siRNA. Representative images of cell migration are shown. (C) Quantification of (B), n = 5, ***p < 0.0003. (D) LR-MSCs transfected with scrambled or FOXF1-specific siRNA were treated with the ATX inhibitor, PF-8380 (1 μM) and migration assay was performed. Values: Means ± SEM. n = 5, **p < 0.0142, *p < 0.0325. (E,F) Migration assays are shown with LR-MSCs transfected with scrambled or FOXF1-specific siRNA, and then treated with the LPA1 inihibitor, VPC12249 (1 μM) (E), or subjected to lentivirus-mediated shRNA interference against LPAR1 (F). Values: Means ± SEM. n = 5. ***p < 0.0002, ****p < 0.0001. Note: Full length blots for Fig. 3A are provided in Supplementary Fig. S5. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/33277571), licensed under a CC-BY license. Not internally tested by R&D Systems.