sirna targeting foxp1  (Santa Cruz Biotechnology)

Journal: Cancer Gene Therapy

Article Title: The FOXP1-ABCG2 axis promotes the proliferation of cancer stem cells and induces chemoresistance in pancreatic cancer

doi: 10.1038/s41417-025-00896-7

Figure Lengend Snippet: A FOXP1 expression levels in PAAD tumor tissues and adjacent normal tissues. FOXP1 is significantly upregulated in tumor tissues. B , C mRNA and protein expression analyses reveal higher FOXP1 expression in PDAC tissue samples than in normal tissues. D Differential gene expression analysis from the GSE71989 dataset reveals high expressions of FOXP1 and CSC markers LEF1 and OCT4 in tumor tissues . E Kaplan–Meier OS and DSS curves for TCGA–pancreatic adenocarcinoma (PAAD) ( F ) mRNA expression levels of FOXP1 and RRM1, a marker of gemcitabine resistance, in GS and GR tissue samples. G FOXP1 and RRM1 protein expression levels in GR tissues. H , I FOXP1 and RRM1 mRNA and protein expression across GS and GR PDAC cell lines, highlighting higher FOXP1 levels in GR cells, with the highest upregulation in Capan-1. * p < 0.05, ** p < 0.01, *** p < 0.0001.

Article Snippet: siRNA-mediated knockdown was performed as previously described [ , ]. siRNA targeting FOXP1 (FOXP1 siRNA, catalog code [sc-44583], Santa Cruz Biotechnology, Dallas, TX, USA) was dissolved and diluted in RNase-free H2O.

Techniques: Expressing, Gene Expression, Marker

Journal: Cancer Gene Therapy

Article Title: The FOXP1-ABCG2 axis promotes the proliferation of cancer stem cells and induces chemoresistance in pancreatic cancer

doi: 10.1038/s41417-025-00896-7

Figure Lengend Snippet: A Flow cytometry (FACS) analysis of CD44+ CD24+ double-positive cells in gemcitabine-sensitive (GS), gemcitabine-resistant (GR), and FOXP1 knockdown (KD) GR Capan-1 cells. ( B ) Colony formation assay comparing GS, GR, and GR FOXP1 KD groups. C Western blot analysis of CSC marker genes in monolayer and 3D spheroid cultures of GS and GR Capan-1 cells. D Spheroid formation over 14 days in GS, GR, and GR FOXP1 KD cells. E Quantitative analysis of mRNA levels of CSC markers in GS, GR, and GR FOXP1 KD cells via qRT-PCR. F Western blot of CSC markers in Capan-1 GS and GR cells with and without FOXP1 knockdown. Data are represented as the mean ± S.D. of three independent experiments ( n = 3). Statistical analysis was conducted using one-way or two-way ANOVA, followed by Tukey’s multiple-comparison test. ** p < 0.01, *** p < 0.0001.

Article Snippet: siRNA-mediated knockdown was performed as previously described [ , ]. siRNA targeting FOXP1 (FOXP1 siRNA, catalog code [sc-44583], Santa Cruz Biotechnology, Dallas, TX, USA) was dissolved and diluted in RNase-free H2O.

Techniques: Flow Cytometry, Knockdown, Colony Assay, Western Blot, Marker, Quantitative RT-PCR, Comparison

Journal: Cancer Gene Therapy

Article Title: The FOXP1-ABCG2 axis promotes the proliferation of cancer stem cells and induces chemoresistance in pancreatic cancer

doi: 10.1038/s41417-025-00896-7

Figure Lengend Snippet: A Wound healing assay of GS, GR, and KD Capan-1 cells at 0, 16, and 24 h. Scale bar = 100 μm ( B ) The number of invaded cells was measured via Transwell Invasion Assay. Scale bar = 100 μm ( C ) Western blot analysis of epithelial and mesenchymal markers in GS, GR, and FOXP1 KD GR cells. D WST-1 cell viability analysis of GS, GR, and FOXP1 KD GR cells. E Immunofluorescence staining of Ki-67 in GS, GR, and FOXP1 KD GR cells. Scale bar = 50 μm ( F ) Western blot analysis of activated MAPK protein expression levels in GS, GR, and FOXP1 KD GR cells. G Western blot analysis of expression levels of cell cycle regulators genes in GS, GR, and FOXP1 KD GR cells. Data are represented as the mean ± S.D. of three independent experiments ( n = 3). Statistical analysis was conducted using one-way or two-way ANOVA, followed by Tukey’s multiple-comparison test. ** p < 0.01, *** p < 0.0001.

Article Snippet: siRNA-mediated knockdown was performed as previously described [ , ]. siRNA targeting FOXP1 (FOXP1 siRNA, catalog code [sc-44583], Santa Cruz Biotechnology, Dallas, TX, USA) was dissolved and diluted in RNase-free H2O.

Techniques: Wound Healing Assay, Transwell Invasion Assay, Western Blot, Immunofluorescence, Staining, Expressing, Comparison

Journal: Cancer Gene Therapy

Article Title: The FOXP1-ABCG2 axis promotes the proliferation of cancer stem cells and induces chemoresistance in pancreatic cancer

doi: 10.1038/s41417-025-00896-7

Figure Lengend Snippet: A GSEA of the TCGA-PAAD cohort shows that high FOXP1 expression is associated with increased glucose metabolism and serum lactate levels. Heatmap analysis of RNA sequencing data from Capan-1 GS and GR cells revealed the upregulation of glycolysis-related genes in GR cells. B Glucose uptake assay showed reduced lactate levels in KD cells compared to GR cells. C mRNA expression analysis of glycolytic genes in GS, GR, and FOXP1 KD GR cells, quantified by qRT-PCR. D Lactate secretion assay showing reduced lactate levels in KD cells compared with GR cells. E mRNA expressions of LDHA and MCT4 in GS, GR, and FOXP1 KD GR cells were quantified by qRT-PCR. F Western blotting analysis of glycolytic enzymes in GS, GR, and KD cells. G OCR and ECAR assays indicated reduced glycolysis in KD cells. Data are represented as the mean ± S.D. of three independent experiments ( n = 3). Statistical analyses were conducted using one-way or two-way ANOVA, followed by Tukey’s multiple-comparison test. * p < 0.05, ** p < 0.01.

Article Snippet: siRNA-mediated knockdown was performed as previously described [ , ]. siRNA targeting FOXP1 (FOXP1 siRNA, catalog code [sc-44583], Santa Cruz Biotechnology, Dallas, TX, USA) was dissolved and diluted in RNase-free H2O.

Techniques: Expressing, RNA Sequencing, Quantitative RT-PCR, Western Blot, Comparison

Journal: Cancer Gene Therapy

Article Title: The FOXP1-ABCG2 axis promotes the proliferation of cancer stem cells and induces chemoresistance in pancreatic cancer

doi: 10.1038/s41417-025-00896-7

Figure Lengend Snippet: A Correlation analysis between FOXP1 and ABCG2 expression levels in the TCGA-PAAD dataset via TIMER 2.0. B Western Blot shows reduced ABCG2 expression in FOXP1 KD cells. C WST-1 viability assay of GR cells treated with KO143 at different concentrations. Co-treatment with gemcitabine and KO143 further decreases cell viability. D Sphere formation assay of GS, GR, FOXP1 KD, KO143-treated GR cells, and co-treatment of KO143 and siFOXP1. E FOXP1 binding motif and schematic of the ABCG2 promoter region showing potential FOXP1 binding sites identified via JASPAR. F Primer design for assessing FOXP1 binding to the ABCG2 promoter. G Luciferase reporter assay in HEK293 cells transfected with wild-type (WT) or mutant (MT) ABCG2 promoter constructs. H Chromatin immunoprecipitation (ChIP) assay in HEK293 cells confirms FOXP1 binding to the ABCG2 promoter. Data are represented as the mean ± S.D. of three independent experiments ( n = 3). Statistical analysis was conducted using one-way or two-way ANOVA, followed by Tukey’s multiple-comparison test. ns > 0.05 , * p < 0.05, ** p < 0.01, *** p < 0.0001.

Article Snippet: siRNA-mediated knockdown was performed as previously described [ , ]. siRNA targeting FOXP1 (FOXP1 siRNA, catalog code [sc-44583], Santa Cruz Biotechnology, Dallas, TX, USA) was dissolved and diluted in RNase-free H2O.

Techniques: Expressing, Western Blot, Viability Assay, Tube Formation Assay, Binding Assay, Luciferase, Reporter Assay, Transfection, Mutagenesis, Construct, Chromatin Immunoprecipitation, Comparison

Journal: Cancer Gene Therapy

Article Title: The FOXP1-ABCG2 axis promotes the proliferation of cancer stem cells and induces chemoresistance in pancreatic cancer

doi: 10.1038/s41417-025-00896-7

Figure Lengend Snippet: A Representative image of tumors from BALB/C nude mice injected with GS or GR Capan-1 cells, with GR cells treated with shFOXP1 lentivirus and/or KO143 (Co-Tx). B Quantification of tumor weights after 5 weeks shows significantly smaller tumors in the Co-Tx group. C Western blot analysis of FOXP1 expressions in GS, GR, shFOXP1-treated, KO143-treated, and Co-Tx groups. D Relative mRNA expression levels of stemness markers show significant reductions in the Co-Tx group compared to untreated groups. E , F Immunofluorescence analyses of FOXP1, ABCG2, and stemness marker genes in tumor tissues from each group. Data are represented as the mean ± S.D. of three independent experiments ( n = 3). Statistical analysis was conducted using one-way or two-way ANOVA, followed by Tukey’s multiple-comparison test. * p < 0.05, ** p < 0.01, *** p < 0.0001.

Article Snippet: siRNA-mediated knockdown was performed as previously described [ , ]. siRNA targeting FOXP1 (FOXP1 siRNA, catalog code [sc-44583], Santa Cruz Biotechnology, Dallas, TX, USA) was dissolved and diluted in RNase-free H2O.

Techniques: Injection, Western Blot, Expressing, Immunofluorescence, Marker, Comparison

Journal: EBioMedicine

Article Title: FOXP1 negatively regulates tumor infiltrating lymphocyte migration in human breast cancer

doi: 10.1016/j.ebiom.2018.11.066

Figure Lengend Snippet: FOXP1 expression in breast cancer cell lines and primary breast cancer. (a) FOXP1 gene and protein expression in breast epithelial tumor cell lines [(MCF7 (ER+/HER2-), BT474 (HER2+) and MDA-MB-231 (triple negative: TN)] by RT-qPCR and immunoblotting. All data were obtained as the mean of three independent experiments. Blots were probed with anti-β actin as subcellular compartment control and isotype control for FOXP1 was also used. (b) Microarray data from METABRIC dataset were analyzed for FOXP1 gene expression (log2) among the breast cancer molecular subtypes ( n = 1992). (c) Evaluation of FOXP1 gene expression using RT-qPCR was performed in a cohort of primary breast tumors ( n = 94) from the tumor banks of Institut Jules Bordet. FOXP1 expression in normal breast also analyzed ( n = 10). RT-qPCR data are relative to EIF1α and MLN51 expression (2 -ΔCt ) done in duplicates shown in log2 scale. Significant P values (<0.05) are marked by *. Degrees of significance: P < 0.05 (*), P < 0.001 (***) and P < 0.0001 (****), as assessed with ANOVA using Dunn's multiple comparisons test. Data represent mean ± SEM.

Article Snippet: The FOXP1 gene was silenced in MCF7 cells using predesigned siRNA-targeted sequences directed against FOXP1 isoforms (Qiagen; SI00421120, SI03196655 and SI04954663).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Control, Microarray, Gene Expression

Journal: EBioMedicine

Article Title: FOXP1 negatively regulates tumor infiltrating lymphocyte migration in human breast cancer

doi: 10.1016/j.ebiom.2018.11.066

Figure Lengend Snippet: FOXP1 protein expression in primary breast cancer. a) Representative FOXP1 staining in normal breast tissue, ER+/HER2-, HER2+ and triple negative breast cancer are shown. FOXP1 staining in stromal cells and TIL in a triple negative is shown in the bottom panel. All images are at magnifications of 200×. (b) The frequency of FOXP1 protein expression in different cell subpopulations and frequency of FOXP1 tumor cells among breast cancer molecular subtypes using IHC. FOXP1 positivity in tumor cells, stromal and TIL have been considered while FOXP1 expression in tumor cells in ER+/HER2-, HER2+ and triple negative were analyzed. FOXP1 positivity was defined as ≥5% of any positive cell among all cells ( n = 104). Significant P values (<0.05) are marked by *. Degrees of significance: P < 0.05 (*) and P < 0.01 (**), as assessed with ANOVA using Dunn's multiple comparisons test. Data represent mean ± SD. IHC: Immunohistochemistry.

Article Snippet: The FOXP1 gene was silenced in MCF7 cells using predesigned siRNA-targeted sequences directed against FOXP1 isoforms (Qiagen; SI00421120, SI03196655 and SI04954663).

Techniques: Expressing, Staining, Immunohistochemistry

Journal: EBioMedicine

Article Title: FOXP1 negatively regulates tumor infiltrating lymphocyte migration in human breast cancer

doi: 10.1016/j.ebiom.2018.11.066

Figure Lengend Snippet: Relationship between FOXP1 and TIL infiltration. Assessment of global TIL, stromal TIL, intra-tumoral TIL, T cell infiltration, B cell infiltration and TLS based on CD3/CD30 IHC in FOXP1lo (n = 49) and FOXP1hi (n = 45) breast cancer. Tumors (n = 94) were divided according to the median value of FOXP1 gene expression. Significant P values (< 0.05) are marked by *. Degrees of significance: P < 0.05 (*), P < 0.01 (**), and P < 0.001 (***), as assessed with Mann Whitney test. Data represent mean ± SD.

Article Snippet: The FOXP1 gene was silenced in MCF7 cells using predesigned siRNA-targeted sequences directed against FOXP1 isoforms (Qiagen; SI00421120, SI03196655 and SI04954663).

Techniques: Gene Expression, MANN-WHITNEY

Journal: EBioMedicine

Article Title: FOXP1 negatively regulates tumor infiltrating lymphocyte migration in human breast cancer

doi: 10.1016/j.ebiom.2018.11.066

Figure Lengend Snippet: FOXP1 is associated with chemokine and cytokine gene expression in breast cancer. (a) Chemokines gene expression, (b) Immuno-modulatory molecules, and (c) immuno-suppressive cytokine gene expression in normal breast tissues (n = 10), FOXP1 lo ( n = 25) and FOXP1 hi (n = 25) breast cancer. RT-qPCR data were normalized using EIF1α and MLN51 expression (2 -ΔCt ). Duplicate samples were analyzed. Significant P values (<0.05) are marked by * and ns: not significant. Degrees of significance: P < 0.05 (*), P < 0.01 (**) and P < 0.001 (***). Data represent mean ± SD.

Article Snippet: The FOXP1 gene was silenced in MCF7 cells using predesigned siRNA-targeted sequences directed against FOXP1 isoforms (Qiagen; SI00421120, SI03196655 and SI04954663).

Techniques: Gene Expression, Quantitative RT-PCR, Expressing

Journal: EBioMedicine

Article Title: FOXP1 negatively regulates tumor infiltrating lymphocyte migration in human breast cancer

doi: 10.1016/j.ebiom.2018.11.066

Figure Lengend Snippet: CXCL13 protein expression in breast cancer. Representative images of CXCL13 protein expression in (a) FOXP1 lo and (b) FOXP1 hi primary breast cancer acquired by confocal microscopy. Yellow scale bars: 50 μm and white scale bars: 100 μm. Contrast was enhanced by turning down DAPI (grey) intensity in merged images. CXCL13+ (blue), CD20+ (green) for B tumor infiltrating lymphocytes and cytokeratin 8/18 (red) for tumor epithelial cells were shown as single channel and merged images.

Article Snippet: The FOXP1 gene was silenced in MCF7 cells using predesigned siRNA-targeted sequences directed against FOXP1 isoforms (Qiagen; SI00421120, SI03196655 and SI04954663).

Techniques: Expressing, Confocal Microscopy

Journal: EBioMedicine

Article Title: FOXP1 negatively regulates tumor infiltrating lymphocyte migration in human breast cancer

doi: 10.1016/j.ebiom.2018.11.066

Figure Lengend Snippet: FOXP1 regulates Lymphocyte migration in breast tumors. Healthy donor PBMC migration in FOXP1 lo and FOXP1 hi tumor supernatants (SN) from primary breast cancer cohort. Top panel shows number of CD45 + cells migrated toward breast cancer SN and fold change. Bottom panel shows fold change of CD4 + T cells (CD3 + CD4 + ); CD8 + T cells (CD3 + CD8 + ) and B cells (CD19 + ) migrated toward BC SN after 24 h of incubation in the transwell assay. Data demonstrated as the mean of biological replicates done in duplicates of FOXP1 lo ( n = 5) and FOXP1 hi (n = 5). Medium alone was used as a negative control and medium with 10% FBS was used a positive control. Significant P values (<0.05) are marked by * and ns: not significant. Degrees of significance: P < 0.05 (*), P < 0.01 (**) and P < 0.001 (***), as assessed with ANOVA using Dunn's multiple comparisons test. Data represent mean ± SEM.

Article Snippet: The FOXP1 gene was silenced in MCF7 cells using predesigned siRNA-targeted sequences directed against FOXP1 isoforms (Qiagen; SI00421120, SI03196655 and SI04954663).

Techniques: Migration, Incubation, Transwell Assay, Negative Control, Positive Control

Journal: EBioMedicine

Article Title: FOXP1 negatively regulates tumor infiltrating lymphocyte migration in human breast cancer

doi: 10.1016/j.ebiom.2018.11.066

Figure Lengend Snippet: FOXP1 is associated with worst survival in breast cancer (a) Univariate analysis for prognostic effect of FOXP1 gene expression in METABRIC dataset. Forest plots for disease specific survival (DSS) and overall survival (OS) based on FOXP1 gene expression in ER+/HER2- ( n = 1394), HER2+ ( n = 244), triple negative (TN) BC ( n = 271) and all breast cancer ( n = 1992). (b) Multivariable model analyzed in the same dataset with adjustments for treatment (yes vs no), age (≤ 50 vs > 50), tumor size (T0, 1, 2 vs T3, 4), nodal status (negative vs positive), histologic grade (1, 2 vs 3), ER status (negative vs positive), and human epidermal growth factor receptor 2 (HER2) status (negative vs positive). P values <.05 are considered as significant. Proportional hazard assumptions were tested for P -values using cox.zph function.

Article Snippet: The FOXP1 gene was silenced in MCF7 cells using predesigned siRNA-targeted sequences directed against FOXP1 isoforms (Qiagen; SI00421120, SI03196655 and SI04954663).

Techniques: Gene Expression