Journal: Journal of Cachexia, Sarcopenia and Muscle

Article Title: FoxP1 is a transcriptional repressor associated with cancer cachexia that induces skeletal muscle wasting and weakness

doi: 10.1002/jcsm.12666

Figure Lengend Snippet: FoxP1 is up‐regulated in multiple models of cancer cachexia in a FoxO1‐dependent manner. (A) Changes in FoxP1 mRNA in mouse skeletal muscle in various experimental models of cancer cachexia, including the subcutaneous murine colon 26 adenocarcinoma (C26) and Lewis lung carcinoma (LLC) models, and the human pancreas‐liver (L3.6pl) xenograft models, in which human L3.6pl cells were injected into the flank or orthotopically (ortho) into the pancreas. Note that independent unpaired two‐tailed t ‐tests were performed for the different cachexia models. (B–F) FOXP1 protein expression in skeletal muscle of C26 tumour‐bearing and non‐tumour‐bearing (Sham) mice. In panel (C), total FOXP1 expression was quantified by summing the signal from FOXP1A, FOXP1B, and FOXP1C and normalizing to total protein. (G) Transfection of rat solei with a constitutively active FoxO1 expression plasmid (ca. FoxO1) is sufficient to increase FoxP1 transcription, in vivo , as highlighted by its ability to increase a luciferase reporter gene driven by the FoxP1 promoter (RLU = relative light unit). (H) FoxO1 knockdown in mouse tibialis anterior , through transduction of muscles with AAV9 vector encoding FoxO1‐shRNA (or scrambled‐shRNA), prevents the C26‐induced upregulation of FoxP1 mRNA. For panels (A)–(G), depending on data distribution, unpaired two‐tailed t ‐tests or Mann–Whitney tests were performed to test for statistical differences between groups. A one‐way analysis of variance (ANOVA) was conducted to test for statistical differences between means in panel (H). Data are reported as mean ± SEM.

Article Snippet: The FoxP1 promoter reporter plasmid (MPRM17622‐PG02) was purchased from Genecopoeia (Rockville, Maryland, USA).

Techniques: Injection, Two Tailed Test, Expressing, Transfection, Plasmid Preparation, In Vivo, Luciferase, Knockdown, Transduction, Muscles, shRNA, MANN-WHITNEY

Journal: Journal of Cachexia, Sarcopenia and Muscle

Article Title: FoxP1 is a transcriptional repressor associated with cancer cachexia that induces skeletal muscle wasting and weakness

doi: 10.1002/jcsm.12666

Figure Lengend Snippet: Inducible, skeletal muscle‐specific FoxP1 over‐expression induces body and skeletal muscle wasting. FoxP1 over‐expression was induced by injecting mice intraperitoneally with tamoxifen for five consecutive days, followed by maintenance on a tamoxifen diet until study endpoint. Mice were euthanized following 6 weeks of treatment, or upon reaching IACUC‐mandated endpoint, based on cachexia development. (A–C) Tamoxifen treatment (started at 9–15 weeks of age) increases FoxP1 mRNA (A) and protein (B–C) expression in CRE+ FoxP1 iSkmTg/Tg mice compared with controls. (D–I) Skeletal muscle‐specific FoxP1 over‐expression induces body wasting (D–F) and skeletal muscle wasting (G–I). TAM = tamoxifen. (J) Representative images of tibialis anterior cross‐sections stained for wheat germ agglutinin (WGA), myosin heavy chain type I (MHCI), and myosin heavy chain type IIA (MHCIIA). CON = genetic controls. (K) Quantification of tibialis anterior muscle fibre cross‐sectional area (CSA) shows a leftward shift in fibre sizes in response to FoxP1 over‐expression. CSA data were binned and fit with a Gaussian least squares regression. Significance was determined by calculating the extra sum‐of‐squares F test. (L–M) Quantification of type IIA (L) and type IIX/B (M) fibre percentage in genetic control (CON) and CRE+ FoxP1 iSkmTg/Tg tibialis anterior . (N–O) Quantification of type IIA (N) and type IIX/B (O) muscle fibre CSA showing type IIX/B fibre atrophy in tibialis anterior muscles over‐expressing FoxP1. Statistical differences were determined by conducting unpaired two‐tailed t ‐tests or Mann–Whitney tests in panels (A) and (L–O), one‐way ANOVA in panel (C) and two‐way ANOVAs in panels (D)–(I). Data shown are from female mice and are reported as mean ± SEM. Note that data from male CRE+ FoxP1 iSkmTg/Tg mice and CRE‐ littermate controls are presented in Figure S3 .

Article Snippet: The FoxP1 promoter reporter plasmid (MPRM17622‐PG02) was purchased from Genecopoeia (Rockville, Maryland, USA).

Techniques: Over Expression, Expressing, Staining, Control, Muscles, Two Tailed Test, MANN-WHITNEY

Journal: Journal of Cachexia, Sarcopenia and Muscle

Article Title: FoxP1 is a transcriptional repressor associated with cancer cachexia that induces skeletal muscle wasting and weakness

doi: 10.1002/jcsm.12666

Figure Lengend Snippet: Skeletal muscles of FoxP1 iSkmTg/Tg exhibit myopathy. (A) Representative images of haematoxylin and eosin (H&E) stained cross‐sections show increased extracellular space, presence of mononucleated cells, and increased number of myofibres with centralized nuclei in muscles from CRE+ FoxP1 iSkmTg/Tg mice compared with CRE‐ littermate controls (CON). (B) Quantification of total number of fibres presenting internalized nuclei in soleus muscle cross‐sections from CRE+ FoxP1 iSkmTg/Tg mice and CRE‐ littermate controls. (C) Representative muscle cross‐sections from female CRE+ FoxP1 iSkmTg/Tg mice and CRE‐ littermate controls stained with mouse immunoglobulin G (IgG) antibodies and wheat germ agglutinin (WGA). (D–F) muscles from female CRE+ FoxP1 iSkmTg/Tg mice show increased presence of intra‐myofibre endogenous IgG (D) and WGA (E) indicative of muscle damage, and increased percent area positive for WGA (F) compared to CRE‐ littermate controls. Data are reported as mean ± SEM. (G) Skeletal muscle cross‐sections from female CRE+ FoxP1 iSkmTg/Tg mice stained with Alizarin Red S show dysregulation of intracellular Ca 2+ compared with CRE‐ littermate controls. (H) Representative images of tibialis anterior cross‐sections stained with periodic acid‐Schiff (PAS) reveal the accumulation of PAS + vacuoles in muscles of CRE+ FoxP1 iSkmTg/Tg mice compared with CRE‐ littermate controls (CON). Depending on data distribution, unpaired two‐tailed t ‐tests or Mann–Whitney tests were performed to test for statistical differences between groups. Data are reported as mean ± SEM. Note that data from male CRE+ FoxP1 iSkmTg/Tg mice and CRE‐ littermate controls are presented in Figure S6 .

Article Snippet: The FoxP1 promoter reporter plasmid (MPRM17622‐PG02) was purchased from Genecopoeia (Rockville, Maryland, USA).

Techniques: Muscles, Staining, Two Tailed Test, MANN-WHITNEY

Journal: Journal of Cachexia, Sarcopenia and Muscle

Article Title: FoxP1 is a transcriptional repressor associated with cancer cachexia that induces skeletal muscle wasting and weakness

doi: 10.1002/jcsm.12666

Figure Lengend Snippet: Inducible, skeletal muscle‐specific FoxP1 over‐expression causes ultra‐structural alterations. Transmission electron microscopy images show ultra‐structural alterations in muscles from female CRE+ FoxP1 iSkmTg/Tg mice compared with CRE‐ littermate controls. C, core; h, hypercontracted sarcomeres; L, lysosome; m. irr., membrane irregularities; N, internalized nucleus; NC, non‐contractile material; nf, necrotioc fibre; V, vacuoles.

Article Snippet: The FoxP1 promoter reporter plasmid (MPRM17622‐PG02) was purchased from Genecopoeia (Rockville, Maryland, USA).

Techniques: Over Expression, Transmission Assay, Electron Microscopy, Muscles, Membrane

Journal: Journal of Cachexia, Sarcopenia and Muscle

Article Title: FoxP1 is a transcriptional repressor associated with cancer cachexia that induces skeletal muscle wasting and weakness

doi: 10.1002/jcsm.12666

Figure Lengend Snippet: Inducible, skeletal muscle‐specific FoxP1 over‐expression causes skeletal muscle weakness. Specific twitch (A) and tetanic (B) forces recorded from diaphragm strips from CRE+ FoxP1 iSkmTg/Tg mice (orange) and CRE‐ littermate controls (blue). Twitch contraction (C) and half relaxation times (D) show slower kinetics in female CRE+ FoxP1 iSkmTg/Tg mice vs. CRE‐ littermate controls. (E–F) Force–frequency relationships obtained from CRE+ FoxP1 iSkmTg/Tg mice and CRE‐ littermate controls indicate that diaphragms from CRE+ FoxP1 iSkmTg/Tg mice show reduced specific forces, and a leftward shift at low stimulation frequencies (F). Twitch (G) and tetanus (H) forces recorded from soleus muscles harvested from CRE+ FoxP1 iSkmTg/Tg mice (orange) and CRE‐ littermate controls. Depending on data distribution, unpaired two‐tailed t ‐tests or Mann–Whitney tests were performed to test for statistical differences between groups in panels (A–D) and (G–H). Statistical differences between means were tested by conducting two‐way mixed ANOVAs for panels (E) and (F). Data are reported as mean ± SEM.

Article Snippet: The FoxP1 promoter reporter plasmid (MPRM17622‐PG02) was purchased from Genecopoeia (Rockville, Maryland, USA).

Techniques: Over Expression, Muscles, Two Tailed Test, MANN-WHITNEY

Journal: Journal of Cachexia, Sarcopenia and Muscle

Article Title: FoxP1 is a transcriptional repressor associated with cancer cachexia that induces skeletal muscle wasting and weakness

doi: 10.1002/jcsm.12666

Figure Lengend Snippet: Acute FoxP1 over‐expression in skeletal muscle leads to dysregulation of skeletal muscle homeostatic pathways. Mouse tibialis anterior muscles were transfected with a FoxP1 plasmid or empty vector and microarray analyses were conducted on pooled samples ( n = 3 per group). (A‐B) IPA enriched diseases and functions that are up‐regulated (A) and down‐regulated (B) in response to acute FoxP1 over‐expression. (C) RT‐qPCR validation for selected genes involved in muscle structural development, function, and maintenance. Depending on data distribution, unpaired two‐tailed t ‐tests or Mann–Whitney tests were performed to test for statistical differences between groups. Data are reported as mean ± SEM.

Article Snippet: The FoxP1 promoter reporter plasmid (MPRM17622‐PG02) was purchased from Genecopoeia (Rockville, Maryland, USA).

Techniques: Over Expression, Muscles, Transfection, Plasmid Preparation, Microarray, Quantitative RT-PCR, Biomarker Discovery, Two Tailed Test, MANN-WHITNEY

Journal: Journal of Cachexia, Sarcopenia and Muscle

Article Title: FoxP1 is a transcriptional repressor associated with cancer cachexia that induces skeletal muscle wasting and weakness

doi: 10.1002/jcsm.12666

Figure Lengend Snippet: Inducible, skeletal muscle‐specific up‐regulation of FoxP1 impedes muscle regeneration following injury. (A–B) FoxP1 (A) and Acta1 (B) mRNA expression measured in proliferating C2C12 myoblasts (Day 0) and following 1, 2, or 5 days of differentiation into myotubes. (C–G) Tibialis anterior muscles of CRE+ FoxP1 iSkmTg/Tg mice and CRE‐ littermate controls (CON) were injured via direct injection of cardiotoxin on the same day as tamoxifen treatments were initiated. Twenty‐four days post‐injury, FoxP1 iSkmTg/Tg mice showed reduced muscle mass (C), and reduced cross‐sectional area (CSA) of regenerating myofibres, as visualized in tibialis anterior muscle cross‐sections stained with H&E (D) and as quantified in (E–G). Cross‐sectional area data were binned, fit with a Gaussian least squares regression and significance was determined by calculating the extra sum‐of‐squares F test. Statistical differences were determined by conducting one‐way ANOVAs in panels (A) and (B), and unpaired two‐tailed t ‐tests or Mann–Whitney tests in panels (C) and (E). Data are reported as mean ± SEM.

Article Snippet: The FoxP1 promoter reporter plasmid (MPRM17622‐PG02) was purchased from Genecopoeia (Rockville, Maryland, USA).

Techniques: Expressing, Muscles, Injection, Staining, Two Tailed Test, MANN-WHITNEY

Journal: Journal of Cachexia, Sarcopenia and Muscle

Article Title: FoxP1 is a transcriptional repressor associated with cancer cachexia that induces skeletal muscle wasting and weakness

doi: 10.1002/jcsm.12666

Figure Lengend Snippet: FoxP1‐dependent induction of body and skeletal muscle wasting occurs through an HDAC‐dependent manner. (A–I) Tamoxifen‐treated CRE+ FoxP1 iSkmTg/Tg mice received daily intraperitoneal (IP) injections of trichostatin A (TSA, a pan HDAC inhibitor). TSA treatment blunted the FoxP1‐dependent loss of body mass (A) and skeletal muscle mass (B–E). Body and muscle masses are expressed as percent of sex‐matched CRE‐ littermate controls (CON). (F–G) Representative images of FoxP1 iSkmTg/Tg muscle cross‐sections stained with haematoxylin & eosin (H&E, F) and wheat germ agglutinin (WGA, G) show that TSA treatment protects against FoxP1‐induced muscle fibre atrophy (H), myopathy including degeneration/regeneration (I), and extracellular matrix (ECM) deposition (J). Data are normalized to CRE‐ littermate controls and reported as mean ± SEM. Cross‐sectional area data were binned, fit with a Gaussian least squares regression and significance was determined by calculating the extra sum‐of‐squares F test. Depending on data distribution, unpaired two‐tailed t ‐tests or Mann–Whitney tests were performed to test for statistical differences between groups in panels (A–E) and (I and J). [Note that data for mice not treated with TSA originate from the same mice as in Figures , , S3 , and S6 ].

Article Snippet: The FoxP1 promoter reporter plasmid (MPRM17622‐PG02) was purchased from Genecopoeia (Rockville, Maryland, USA).

Techniques: Staining, Two Tailed Test, MANN-WHITNEY

Journal: Journal of Cachexia, Sarcopenia and Muscle

Article Title: FoxP1 is a transcriptional repressor associated with cancer cachexia that induces skeletal muscle wasting and weakness

doi: 10.1002/jcsm.12666

Figure Lengend Snippet: FoxP1 mediates cancer‐induced muscle atrophy and the repression of MEF2 target genes. (A) Transduction of tibialis anterior muscles of C26‐tumour bearing mice with an AAV9 vector encoding FoxP1‐shRNA blunts the C26‐induced reduction in muscle fibre cross‐sectional area (CSA) that is observed in muscles transduced with AAV9 encoding scrambled‐shRNA. (B) Representative transmission electron microscopy images of tibialis anterior muscles from sham and C26‐tumour bearing mice transduced with the AAV9 encoding FoxP1 shRNA or scrambled shRNA, showing ultrastructural alterations in response to C26 tumour burden, that are improved in response to FoxP1 knockdown. (C) Microarray analyses conducted on pooled samples ( n = 3 per group) and further analysed with string to identify enriched cellular components, indicate that FoxP1 knockdown attenuates the tumour‐induced repression of genes involved in muscle structure/function (blue categories) and reduces the activation of genes involved in protein breakdown (red categories). (D–E) Genes repressed by two‐fold or more in response to C26 tumour burden that were rescued by two‐fold or more in muscles expressing FoxP1‐shRNA (249 genes) were further analysed using DAVID to identify enriched non‐redundant functional annotations (D), and using gene set enrichment analysis (GSEA) and the molecular signatures database to identify enriched transcription factor binding motifs among gene promoters (E). FoxP1 targets repressed in muscle of C26 mice were enriched for MEF2 and MYOD target genes involved in muscle contraction and muscle cell differentiation. (F) RT‐qPCR validation demonstrating that FoxP1 knockdown rescues the C26‐induced repression of several MEF2 target genes involved in muscle structural development, function, and maintenance. (G) Mouse tibialis anterior muscles were transfected with a FoxP1 expression plasmid or empty vector, plus a MEF2‐dependent luciferase reporter to assess MEF2 activity. RLU = relative light unit. Statistical differences were determined by conducting a one‐way ANOVA in panel (A), unpaired two‐tailed t ‐test or Mann–Whitney tests depending on data distribution in panel (F) and Wilcoxon test in panel (G). Data are reported as mean ± SEM.

Article Snippet: The FoxP1 promoter reporter plasmid (MPRM17622‐PG02) was purchased from Genecopoeia (Rockville, Maryland, USA).

Techniques: Transduction, Muscles, Plasmid Preparation, shRNA, Transmission Assay, Electron Microscopy, Knockdown, Microarray, Activation Assay, Expressing, Functional Assay, Binding Assay, Cell Differentiation, Quantitative RT-PCR, Biomarker Discovery, Transfection, Luciferase, Activity Assay, Two Tailed Test, MANN-WHITNEY

Journal: Journal of Cachexia, Sarcopenia and Muscle

Article Title: FoxP1 is a transcriptional repressor associated with cancer cachexia that induces skeletal muscle wasting and weakness

doi: 10.1002/jcsm.12666

Figure Lengend Snippet: FoxP1 expression is increased in skeletal muscle of patients with pancreatic ductal adenocarcinoma (PDAC) exhibiting cachexia. (A–B) FOXP1 protein expression is increased in rectus abdominis muscle biopsies from cachectic PDAC patients compared with non‐cancer control patients. In panel (A), arrows point to the two FoxP1 isoforms quantified in B. (C–E) FOXP1 mRNA expression in rectus abdominis muscle of PDAC patients correlates positively with body mass loss (C) and negatively with skeletal muscle index (D) and is increased in cachectic PDAC patients when compared with weight‐stable non‐cancer controls with normal muscularity (E). Note: mRNA data were extracted from microarray analyses previously analysed and published in combination with histological assessment of muscle biopsies ²⁰ and in combination with MRI‐based measures of skeletal muscularity and muscle quality. ⁶⁸ Statistical differences were determined by conducting unpaired two‐tailed t ‐test or Mann–Whitney tests depending on data distribution in panels (B) and (E), and simple linear regression analyses in panels (C) and (D). Data are reported as mean ± SEM.

Article Snippet: The FoxP1 promoter reporter plasmid (MPRM17622‐PG02) was purchased from Genecopoeia (Rockville, Maryland, USA).

Techniques: Expressing, Control, Microarray, Two Tailed Test, MANN-WHITNEY

Journal: Scientific reports

Article Title: LncRNA HOXA-AS3 promotes cell proliferation and invasion via targeting miR-218-5p/FOXP1 axis in osteosarcoma.

doi: 10.1038/s41598-024-67596-4

Figure Lengend Snippet: Figure 3. LncRNA HOXA-AS3 interacts with miR-218-5p in osteosarcoma cells. (A) Potential binding sites between miR-218-5p and lncRNA HOXA-AS3 and FOXP1. (B) RT-PCR data showed that miR-218-5p inhibitors decreased the expression of miR-218-5p. In addition, miR-218-5p mimic transfection increased the miR-218-5p expression levels. (C) Dual luciferase reporter assays showed that miR-218-5p interacted with lncRNA HOXA-AS3. miR-218-5p regulated luciferase activity in HOXA-AS3 wild-type group. (D) Dual luciferase reporter assays showed that miR-218-5p interacted with and regulated FOXP1. **p < 0.01 versus control group. (E) Western blotting analysis data showed that lncRNA HOXA-AS3 and miR-218-5p regulate the expression of FOXP1 in osteosarcoma cells. WT: wild type; MUT: mutant.

Article Snippet: The primary antibody FOXP1 (1:2000, #2005, Cell Signaling Technology) were used to incubate the membrane at cold room overnight.

Techniques: Binding Assay, Reverse Transcription Polymerase Chain Reaction, Expressing, Transfection, Luciferase, Activity Assay, Control, Western Blot, Mutagenesis

Journal: Scientific reports

Article Title: LncRNA HOXA-AS3 promotes cell proliferation and invasion via targeting miR-218-5p/FOXP1 axis in osteosarcoma.

doi: 10.1038/s41598-024-67596-4

Figure Lengend Snippet: Figure 4. Downregulation of lncRNA HOXA-AS3 reduces colony formation via miR-218-5p and FOXP1. (A) Colony formation assays were performed to measure the colony formation in osteosarcoma cells co-transfected with sh-HOXA-AS3 and pcDNA FOXP1, or sh-HOXA-AS3 and miR-218 inhibitors. (B) Quantitative data are shown for colony formation. ***p < 0.001 versus control group. #p < 0.05; ##p < 0.01 versus sh-HOXA-AS3 alone.

Article Snippet: The primary antibody FOXP1 (1:2000, #2005, Cell Signaling Technology) were used to incubate the membrane at cold room overnight.

Techniques: Transfection, Control

Journal: Scientific reports

Article Title: LncRNA HOXA-AS3 promotes cell proliferation and invasion via targeting miR-218-5p/FOXP1 axis in osteosarcoma.

doi: 10.1038/s41598-024-67596-4

Figure Lengend Snippet: Figure 5. Downregulation of lncRNA HOXA-AS3 reduces cell migration via miR-218-5p and FOXP1. (A) Wound healing assays were performed to measure migratory ability in osteosarcoma cells co-transfected with sh-HOXA-AS3 and pcDNA FOXP1, or sh-HOXA-AS3 and miR-218 inhibitors. (B) Quantitative data are shown for colony formation. **p < 0.01; ***p < 0.001 versus control group. #p < 0.05; ##p < 0.01 versus sh-HOXA-AS3 alone.

Article Snippet: The primary antibody FOXP1 (1:2000, #2005, Cell Signaling Technology) were used to incubate the membrane at cold room overnight.

Techniques: Migration, Transfection, Control

Journal: Scientific reports

Article Title: LncRNA HOXA-AS3 promotes cell proliferation and invasion via targeting miR-218-5p/FOXP1 axis in osteosarcoma.

doi: 10.1038/s41598-024-67596-4

Figure Lengend Snippet: Figure 6. Downregulation of lncRNA HOXA-AS3 reduces cell invasion via miR-218-5p and FOXP1. (A) Transwell assays were performed to measure cell invasive ability in osteosarcoma cells co-transfected with sh-HOXA-AS3 and pcDNA FOXP1, or sh-HOXA-AS3 and miR-218 inhibitors. (B) Quantitative data are shown for colony formation. ***p < 0.001 versus control group. #p < 0.05; ##p < 0.01 versus sh-HOXA-AS3 alone.

Article Snippet: The primary antibody FOXP1 (1:2000, #2005, Cell Signaling Technology) were used to incubate the membrane at cold room overnight.

Techniques: Transfection, Control

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: Fig. 1 FOXP1 is upregulated in gemcitabine-resistant (GR) PDAC. 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 [21]. 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 [2, 7]. 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: Fig. 2 FOXP1 promotes the proliferation of CSCs in GR PDAC. 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 [2, 7]. 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: Fig. 4 FOXP1 induces metabolic reprogramming by promoting glycolysis. 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 [2, 7]. 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: Fig. 3 FOXP1 enhances EMT and induces proliferation in GR PDAC. 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 [2, 7]. 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, 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: Fig. 5 FOXP1 chemosensitizes PDAC to gemcitabine by upregulating ABCG2. 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 [2, 7]. 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: Fig. 6 Reduced FOXP1 and ABCG2 expressions lead to reduced tumor growth and increased chemosensitivity in mouse models. 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 [2, 7]. 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, Marker, Comparison

Journal: Nature immunology

Article Title: Genome-wide DNA methylation landscape defines specialization of regulatory T cells in tissues

doi: 10.1038/ni.3799

Figure Lengend Snippet:

Article Snippet: Taqman Probe for Foxp1 , Thermo Fisher , Mm00474848_m1.

Techniques: Affinity Purification, Derivative Assay, SYBR Green Assay, Gene Expression, Multiplex Assay, Reverse Transcription, Functional Assay, Purification, Recombinant, DNA Methylation Assay, Gel Extraction, Staining