primary antibodies against foxm1  (Danaher Inc)

Journal: American Journal of Cancer Research

Article Title: Exosomal miR-664a-5p as a therapeutic target biomarker for PARP inhibitor response in prostate cancer

doi: 10.62347/qyzs2620

Figure Lengend Snippet: Figure 4. MiR-664a-5p directly targets to FOXM1. (A) Schematic structure of the dual luciferase reporter vector, and binding sites between miR-664a-5p and FOXM1 predicted through miRWalk. (B) PC-3 cells were co-transfected with FOXM1 3’UTR-luciferase reporter, and miR-664a-5p mimic or negative control miRNA mimic (miR-NC). After 48 h, relative luciferase activity of the reporter was evaluated by dual luciferase reporter analysis. 3’UTR: 3’ untranslated region. (C, D) Three prostate cancer cells (C4-2B, PC-3, and 22Rv1) were transiently transfected with miR-664a-5p mimic or miR-NC mimic. After 3 days of transfection, FOXM1 mRNA (C) and protein (D) expression was determined by RT-qPCR and western blot analyses, respectively. β-actin was the loading control. **P<0.01, ***P<0.001. Data in (B and C) were presented as the mean ± SD of three experiments performed in triplicate.

Article Snippet: The membranes were blocked and subsequently incubated overnight at 4°C with specific primary antibodies against FOXM1 (1:1000, Santa Cruz Biotechnology, Inc., Dallas, TX, USA) and β-actin (1:5000, Abcam, Cambridge, UK).

Techniques: Luciferase, Plasmid Preparation, Binding Assay, Transfection, Negative Control, Activity Assay, Expressing, Quantitative RT-PCR, Western Blot, Control

Journal: Genes & Diseases

Article Title: Down-regulation of microRNA-23a promotes pancreatic ductal adenocarcinoma initiation and progression by up-regulation of FOXM1 expression

doi: 10.1016/j.gendis.2023.101203

Figure Lengend Snippet: Induction of ADM decreased the expression of miR-23a in primary acinar cells and acinar cells in vitro . (A) Primary acinar cells were treated with TGF-α for three days, immunofluorescence staining was performed using specific antibodies against CK19 and amylase. Note that the induction of ADM was accompanied by an increased level of CK19 and decreased expression of amylase. (B) Primary acinar cells, MPC-83, and 266-6 cells were treated with or without 50 ng/mL TGF-α for three days. miR-23a-3p and miR-23a-5p levels were determined with quantitative PCR in triplicate. Note that induction of ADM correlated with a significantly decreased expression of miR-23a-3p and miR-23a-5p in primary acinar cells and MPC-83 and 266-6 pancreatic cells. ADM, acinar-to-ductal metaplasia.

Article Snippet: Whole-cell lysates were subjected to standard western blot analyses using primary antibodies against FOXM1, pancreatic amylase, and CK19 (Beyotime, CAT# P0013B).

Techniques: Expressing, In Vitro, Immunofluorescence, Staining, Real-time Polymerase Chain Reaction

Journal: Genes & Diseases

Article Title: Down-regulation of microRNA-23a promotes pancreatic ductal adenocarcinoma initiation and progression by up-regulation of FOXM1 expression

doi: 10.1016/j.gendis.2023.101203

Figure Lengend Snippet: Increased FOXM1 expression in ADM correlated with increased fibrosis in the pancreas. Three mouse models of ADM were generated as described in the Materials and Methods section, including PDL, CAE, and KC mouse models. (A) Mouse pancreatic tissue sections were prepared and subjected to immunohistochemical staining utilizing specific antibodies against FoxM1, CK19, amylase, α-SMA, and desmin. Relative expressions of those proteins were determined. The expression level of CK19, α-SMA, desmin, and FoxM1 increased, and the expression level of amylase decreased in PDL, CAE, and KC mouse models compared with wild-type (WT) mouse model ( n = 5). (B) Mouse pancreatic tissue samples were sectioned and subjected to immunofluorescence staining with specific anti-FoxM1, anti-CK19, anti-amylase, anti-α-SMA, and anti-desmin antibodies. PDL, pancreatic ductal ligation; CAE, caerulein treatment; KC, Pdx1-Cre;LSL-Kras G12D/+ ; ADM, acinar-to-ductal metaplasia.

Article Snippet: Whole-cell lysates were subjected to standard western blot analyses using primary antibodies against FOXM1, pancreatic amylase, and CK19 (Beyotime, CAT# P0013B).

Techniques: Expressing, Generated, Immunohistochemical staining, Staining, Immunofluorescence, Ligation

Journal: PeerJ

Article Title: High-risk histological subtype-related FAM83A hijacked FOXM1 transcriptional regulation to promote malignant progression in lung adenocarcinoma

doi: 10.7717/peerj.16306

Figure Lengend Snippet: (A) The correlation of FAM83A and FOXM1, CDC6, CDC20 and CDKN3 in TCGA-LUAD database. (B) Upper: Schematic representation of the Flag and FAM83A fusion plasmid. Bottom: ChIP-PCR of FAM83A in promoter regions of FOXM1. (C) Left: Schematic representation of the FOXM1 promoter region truncated luciferase reporter plasmids. Right: Luciferase reporter gene assay for FOXM1 promoter region truncated plasmids. (D) Immunofluorescence assay for FOXM1 in A549 cells with overexpression FAM83A. (E) The expression of FOXM1 was detected after nuclear cytoplasmic separation assay in A549 cells with overexpression FAM83A. * p < 0.05, ** p < 0.01, *** p < 0.001 (Student’s t test).

Article Snippet: Primary antibodies against FOXM1 (ab207298, dilution: 1/1,000), FAM83A (ab128245, dilution: 1/1,000), CDC20 (ab183479, dilution: 1/2,000), CDC6 (ab109315, dilution: 1/4,000), cyclin B (ab32053, dilution: 1/8,000), H3 (ab1791, dilution: 1/5,000), and GAPDH (ab9485, dilution: 1/2,500) were bought from Abcam for this work.

Techniques: Plasmid Preparation, Luciferase, Reporter Gene Assay, Immunofluorescence, Over Expression, Expressing

Journal: PeerJ

Article Title: High-risk histological subtype-related FAM83A hijacked FOXM1 transcriptional regulation to promote malignant progression in lung adenocarcinoma

doi: 10.7717/peerj.16306

Figure Lengend Snippet: Colony formation (A) and cell cycle assays (B) revealed that cell proliferation affected by FAM83A knockdown or overexpression was reversed by cotransfection with FOXM1 knockdown or overexpression. All the results were shown as mean ± SD ( n = 3), which were three separate experiments performed in triplicate. ** p < 0.01, *** p < 0.001 (Student’s t test). Colony formation (C) and cell cycle assays (D) revealed that the efficacy of Palbociclib was more significant after FAM83A overexpression.

Article Snippet: Primary antibodies against FOXM1 (ab207298, dilution: 1/1,000), FAM83A (ab128245, dilution: 1/1,000), CDC20 (ab183479, dilution: 1/2,000), CDC6 (ab109315, dilution: 1/4,000), cyclin B (ab32053, dilution: 1/8,000), H3 (ab1791, dilution: 1/5,000), and GAPDH (ab9485, dilution: 1/2,500) were bought from Abcam for this work.

Techniques: Over Expression, Cotransfection

Journal: PeerJ

Article Title: High-risk histological subtype-related FAM83A hijacked FOXM1 transcriptional regulation to promote malignant progression in lung adenocarcinoma

doi: 10.7717/peerj.16306

Figure Lengend Snippet: (A) Representative images of H&E staining, anti-ki67, anti-FAM83A and anti-FOXM1 immunohistochemistry in 12 LUAD patients. (B) According to the FAM83A IHC score of 12 LUAD patients, they were divided into high expression group and low expression group. (C) The IHC scores of Ki67 and FOXM1 were compared between the FAM83A high expression group and the FAM83A low expression group. (D) The overall survival analysis for patients with different expression of FAM83A and FOXM1 in TCGA-LUAD database. (E) The expression of cell cycle-related biomarker genes in patients with different expression of FAM83A and FOXM1 in TCGA-LUAD database. *** p < 0.001 (Student’s t test).

Article Snippet: Primary antibodies against FOXM1 (ab207298, dilution: 1/1,000), FAM83A (ab128245, dilution: 1/1,000), CDC20 (ab183479, dilution: 1/2,000), CDC6 (ab109315, dilution: 1/4,000), cyclin B (ab32053, dilution: 1/8,000), H3 (ab1791, dilution: 1/5,000), and GAPDH (ab9485, dilution: 1/2,500) were bought from Abcam for this work.

Techniques: Staining, Immunohistochemistry, Expressing, Biomarker Assay