Human FoxM1 Antibody

FoxM1 in U2OS Human Cell Line. FoxM1 was detected in immersion fixed U2OS human osteosarcoma cell line using 10 µg/mL Sheep Anti-Human FoxM1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3975) for 3 hours at room temperature. Cells were stained with the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; Catalog # NL010) and counterstained (green). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Detection of Human FoxM1 by Western Blot. Western blot shows lysates of U2OS human osteosarcoma cell line. PVDF membrane was probed with 1 µg/mL of Sheep Anti-Human FoxM1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3975) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for FoxM1 at approximately 100 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.

Detection of Human FoxM1 by Western Blot RASSF1A regulation is dependent on FoxM1. T84 and Colo 205 cells were transfected with plasmid for overexpression of FoxM1. (A) Cell lysates were analyzed by immunoblot and quantified by densitometry for expression of FoxM1, RASSF1A. Expression is normalized against GAPDH. The right panel of (A) represents the densitometric analysis of FoxM1 and RASSF1A. (B) T84 and Colo 205 cells were transfected with siRNA for FoxM1 or control siRNA for 48 h. Cell lysates were evaluated for FoxM1 ((B), 1st lane), RASSF1A ((B), 2nd lane) by immunoblot and quantified by densitometry. The results are from three independent experiments. (** p < 0.01, *** p < 0.001). Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30744076), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human FoxM1 by Western Blot Neutralizing VEGF receptor antibodies and Akt inhibition upregulates RASSF1A and downregulates FoxM1 in mCRC. T84 and Colo 205 cells were treated with VEGF receptor 1 and VEGFR2 as indicated before (see Section 2.3). Protein expression from cell lysates was determined by immunoblotting for RASSF1A ((A), upper panel) and FoxM1 ((A), middle panel). Densitometry analysis was performed and normalyzed with GAPDH expression to demonstrate significant upregulation for RASSF1A (B) and downregulation of FoxM1 in the presence of neutralized (NT) VEGFR1 or VEGFR2 (C). (D,E) T84 and Colo 205 cells were treated with Akt inhibitor (wortmannin) with different doses (0–1 µM) for 24 h. Cell lysates were analyzed for pAkt (1st lane), total Akt (2nd lane), RASSF1A (3rd lane) expression by immunoblot analysis and quantified by densitometry (F,G). The results are from three independent experiments. (* p < 0.05, ** p < 0.01, *** p < 0.001). Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30744076), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human FoxM1 by Western Blot RASSF1A regulation is dependent on FoxM1. T84 and Colo 205 cells were transfected with plasmid for overexpression of FoxM1. (A) Cell lysates were analyzed by immunoblot and quantified by densitometry for expression of FoxM1, RASSF1A. Expression is normalized against GAPDH. The right panel of (A) represents the densitometric analysis of FoxM1 and RASSF1A. (B) T84 and Colo 205 cells were transfected with siRNA for FoxM1 or control siRNA for 48 h. Cell lysates were evaluated for FoxM1 ((B), 1st lane), RASSF1A ((B), 2nd lane) by immunoblot and quantified by densitometry. The results are from three independent experiments. (** p < 0.01, *** p < 0.001). Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30744076), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human FoxM1 by Western Blot FoxM1 and RASSF1A expression is inversely co-related in colon cancer tissues. (A) Tissue extranction from both normal and colon cancer patient for monitoring the translational level of FoxM1 and RASSF1A by immunoblot. Quantification of RASSF1A (B) and FoxM1 (C) by scanning densitometry. GAPDH was used as a loading control. (D) T84 and Colo 205 cells were treated with or without AGP IC50 (45 µM) for 48 h. Cell lysates were analyzed by Western blot for FoxM1 and GAPDH expression. (E) Quantitative estimations of FoxM1 levels determined by densitometry measurements of western blots from three independent experiments after normalization with GAPDH (p < 0.001). T84 and Colo 205 cells were treated with or without AGP as indicated before and the transcriptional level were determined by qRT-PCR for (F) FoxM1 and (G) PTTG1. Bar graphs show quantitative results normalized to GAPDH mRNA levels. Results are from three independent experiments and statistical significance was determined using one way-ANOVA followed Bonferroni test. (* p < 0.05, ** p < 0.01, *** p < 0.001). Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30744076), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human FoxM1 by Western Blot FoxM1 inhibitor induced RASSF1A and PTEN expression and YAP phosphorylation in mCRC cells. T84 and Colo 205 cells were treated with FoxM1 inhibitor, thiostrepton (Th) with different doses (0–8 µM) for 48 h. (A,B) mRNA was extracted after 24 h for detection of RASSF1A by qRT-PCR. Bar graphs show quantitative results normalized to GAPDH mRNA levels. Cell lysates were monitored by immunoblot for FoxM1(C), RASSF1A (D), p-YAP and total YAP (E), PTEN (F), and GAPDH as indicated. Immunoblots were quantified by scanning densitometry and normalized against GAPDH expression (lower panels of (C–E) for FoxM1, RASSF1A and p-YAP, respectively. Right panel of (F) represents the quantification of PTEN). (G) Patient derived organoids were treated with (8 µM) and without thiostrepton for 24 h and 48 h. Organoid cell lysates were analyzed by immunoblot for RASSF1A expression and quantified by densitometry ((G), lower panel). The results are from three independent experiments. (* p < 0.05, ** p < 0.01, *** p < 0.001). Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30744076), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human FoxM1 by Western Blot FoxM1 and RASSF1A expression is inversely co-related in colon cancer tissues. (A) Tissue extranction from both normal and colon cancer patient for monitoring the translational level of FoxM1 and RASSF1A by immunoblot. Quantification of RASSF1A (B) and FoxM1 (C) by scanning densitometry. GAPDH was used as a loading control. (D) T84 and Colo 205 cells were treated with or without AGP IC50 (45 µM) for 48 h. Cell lysates were analyzed by Western blot for FoxM1 and GAPDH expression. (E) Quantitative estimations of FoxM1 levels determined by densitometry measurements of western blots from three independent experiments after normalization with GAPDH (p < 0.001). T84 and Colo 205 cells were treated with or without AGP as indicated before and the transcriptional level were determined by qRT-PCR for (F) FoxM1 and (G) PTTG1. Bar graphs show quantitative results normalized to GAPDH mRNA levels. Results are from three independent experiments and statistical significance was determined using one way-ANOVA followed Bonferroni test. (* p < 0.05, ** p < 0.01, *** p < 0.001). Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30744076), licensed under a CC-BY license. Not internally tested by R&D Systems.