Journal: Frontiers in Pharmacology

Article Title: Profilin 1 Induces Tumor Metastasis by Promoting Microvesicle Secretion Through the ROCK 1/p-MLC Pathway in Non-Small Cell Lung Cancer

doi: 10.3389/fphar.2022.890891

Figure Lengend Snippet: PFN1 is correlated with NSCLC metastasis and could promote NSCLC cell migration in vitro . (A) Representative IHC images of PFN1 expression on the NSCLC tissues. (B) The staining index of PFN1 on the tissue chip. ** p < 0.01. (C) Representative IHC images of PFN1 expression on the tissue chip. (D) The expression of PFN1 in TCGA LUAD data. ** p < 0.01. (E) The Kaplan–Meier survival analysis of PFN1 in NSCLC patients. (Data source: TCGA LUAD dataset) (F,G) Wound healing assays conducted to evaluate the migration ability of PFN1 -overexpressing (F) and PFN1 knockdown (KD) (G) H1299 cells. ** p < 0.01; scale bar, 500 μm. (H,I) Transwell migration assays conducted to evaluate the migration of PFN1 -overexpressing (H) and PFN1 KD (I) H1299 cells. ** p < 0.01; scale bar, 500 μm. EV, empty vector; OE, PFN1 overexpression; NC, negative control; si-1/ 2, PFN1 siRNA1 1/2.

Article Snippet: PFN1 siRNA and control scramble siRNA were synthesized by Guangzhou RiboBio Co. (Guangzhou, China), and the sequences are listed in .

Techniques: Migration, In Vitro, Expressing, Staining, Knockdown, Plasmid Preparation, Over Expression, Negative Control

Journal: Frontiers in Pharmacology

Article Title: Profilin 1 Induces Tumor Metastasis by Promoting Microvesicle Secretion Through the ROCK 1/p-MLC Pathway in Non-Small Cell Lung Cancer

doi: 10.3389/fphar.2022.890891

Figure Lengend Snippet: PFN1 could promote MVs secretion in NSCLC. (A) Heatmap of differentially expressed proteins between EV and PFN1 OE cells. (B) GO enrichment analysis of differentially expressed proteins. (C) COG/KOG analysis of differentially expressed proteins. (D) MVs extracted from EV-expressing and PFN1 -overexpressing cells, using continuous differential centrifugation, identified using transmission electron microscopy. Scale bar, 100 nm. (E,F) Flow cytometry (E) and western blotting (F) were used to quantify MVs in PFN1 -overexpressing and EV-expressing cells. ARF6 and actin were used as MV markers. (G) Expression of PFN1 and annexin A1 in lung tumor tissues detected using immunofluorescence. (H) The staining index of p-MLC on the tissue chip. ** p < 0.01. (I) Representative IHC images of p-MLC expression. (J) Spearman rank correlation analysis was used to assess the relationship between PFN1 and p-MLC expression on the tissue chip; p and r values are shown in the plot.

Article Snippet: PFN1 siRNA and control scramble siRNA were synthesized by Guangzhou RiboBio Co. (Guangzhou, China), and the sequences are listed in .

Techniques: Expressing, Centrifugation, Transmission Assay, Electron Microscopy, Flow Cytometry, Western Blot, Immunofluorescence, Staining

Journal: Frontiers in Pharmacology

Article Title: Profilin 1 Induces Tumor Metastasis by Promoting Microvesicle Secretion Through the ROCK 1/p-MLC Pathway in Non-Small Cell Lung Cancer

doi: 10.3389/fphar.2022.890891

Figure Lengend Snippet: MVs derived from PFN1 OE cells promote migration in NSCLC cells. (A) MVs collected from sera of patients with NSCLC quantified using flow cytometry. ** p < 0.01. (B) Protein expression of ARF6 and β-actin in MVs collected from sera of patients with NSCLC detected using western blotting. (C) Effect of PFN1 -overexpressing cell supernatants on cell migration evaluated through wound healing assays. ** p < 0.01; scale bar, 500 μm. (D) PKH67-labeled MVs taken up by H1299 cells. DAPI was used to stain the nuclei of H1299 cells. Scale bar, 500 μm. (E,F) Wound healing (E) and Transwell migration (F) assays conducted to evaluate the migration of H1299 cells after treatment with MVs derived from EV-expressing and PFN1 -overexpressing cells; ** p < 0.01; scale bar, 500 μm.

Article Snippet: PFN1 siRNA and control scramble siRNA were synthesized by Guangzhou RiboBio Co. (Guangzhou, China), and the sequences are listed in .

Techniques: Derivative Assay, Migration, Flow Cytometry, Expressing, Western Blot, Labeling, Staining

Journal: Frontiers in Pharmacology

Article Title: Profilin 1 Induces Tumor Metastasis by Promoting Microvesicle Secretion Through the ROCK 1/p-MLC Pathway in Non-Small Cell Lung Cancer

doi: 10.3389/fphar.2022.890891

Figure Lengend Snippet: PFN1 promotes in vivo NSCLC metastasis by elevating MV secretion. (A) Schematic illustration of the mouse model of metastatic tumor established to determine the role of PFN1 in tumor metastasis. (B) Body weight changes in mice after intracardiac injection of PFN1 -overexpressing and EV-expressing cell lines. (C,D) Representative images of lung (C) and liver (D) metastases of the mouse model. The number of metastases is displayed in the right-hand side graph. * p < 0.05, ** p < 0.01. (E) Representative images of HE-stained lung tissues of the mouse model. (F) Representative IHC images of PFN1 and p-MLC expression in lung tissues. The staining index is shown in the right-hand side graph. ** p < 0.01. (G) Representative images of HE-stained liver tissues of the mouse model. (H) Representative IHC images of PFN1 and p-MLC expression in liver tissues. The staining index is shown in the right-hand side graph. ** p < 0.01. (I) Body weight changes in mice after intracardiac injection of H1299 cells and MVs. (J) Representative images of lung metastases of the mouse model. The number of metastases is shown in the bottom graph. * p < 0.05. (K) Representative images of HE-stained lung tissues of the mouse model. (L) Representative IHC images of PFN1 and p-MLC expression in lung tissues. The staining index is shown in the right-hand side graph; * p < 0.05.

Article Snippet: PFN1 siRNA and control scramble siRNA were synthesized by Guangzhou RiboBio Co. (Guangzhou, China), and the sequences are listed in .

Techniques: In Vivo, Injection, Expressing, Staining

Journal: Frontiers in Pharmacology

Article Title: Profilin 1 Induces Tumor Metastasis by Promoting Microvesicle Secretion Through the ROCK 1/p-MLC Pathway in Non-Small Cell Lung Cancer

doi: 10.3389/fphar.2022.890891

Figure Lengend Snippet: Mechanisms underlying the promotion of MLC phosphorylation by PFN1. (A,B) Protein expression after PFN1 overexpression (A) and knockdown (B) measured using western blotting. (C) Protein expression in PFN1 mutants measured using western blotting. (D) PFN1 interactions with ROCK1/2 confirmed using co-IP. (E) Protein expression after treatment with Y27632 (10 µM) measured using western blotting. (F) Effect of PFN1 on ROCK1 activity. ** p < 0.01. (G) Effect of PFN1 on ROCK2 activity. (H) Flow cytometry measuring changes in the amount of MVs after treatment with Y27632; * p < 0.05.

Article Snippet: PFN1 siRNA and control scramble siRNA were synthesized by Guangzhou RiboBio Co. (Guangzhou, China), and the sequences are listed in .

Techniques: Phospho-proteomics, Expressing, Over Expression, Knockdown, Western Blot, Co-Immunoprecipitation Assay, Activity Assay, Flow Cytometry

Journal: Frontiers in Pharmacology

Article Title: Profilin 1 Induces Tumor Metastasis by Promoting Microvesicle Secretion Through the ROCK 1/p-MLC Pathway in Non-Small Cell Lung Cancer

doi: 10.3389/fphar.2022.890891

Figure Lengend Snippet: ROCK1 inhibitor Y27632 partially reversed the promotion of lung cancer metastasis by PFN1 in vitro and in vivo . (A,B) Wound healing assays conducted to evaluate the effect of Y27632 (A) and Y27632 combined with MVs (B) on cell migration. ** p < 0.01; scale bar, 500 μm. (C) Transwell migration assays conducted to evaluate the effect of Y27632 and Y27632 combined with MVs on cell migration. ** p < 0.01; scale bar, 500 μm. (D) Schematic diagram of the mouse model of metastatic tumor established to determine the effect of Y27632 on PFN1-induced lung cancer metastasis. (E) Body weight changes in mice after intracardiac injection of PFN1 -overexpressing H1299 cells and intraperitoneal injection of Y27632 (10 mg/kg). (F) Representative images of lung and liver metastatic tissue in mice. The number of metastatic nodules is shown in the right-hand side graph. * p < 0.05. (G,H) Representative images of HE-stained lung (G) and liver (H) metastases. (I) Representative IHC images of PFN1 and p-MLC expression in lung tissues. The staining index is shown in the right-hand side graph. ** p < 0.01. (J) Representative IHC images of PFN1 and p-MLC expression in liver tissues. The staining index is shown in the right-hand side graph; ** p < 0.01.

Article Snippet: PFN1 siRNA and control scramble siRNA were synthesized by Guangzhou RiboBio Co. (Guangzhou, China), and the sequences are listed in .

Techniques: In Vitro, In Vivo, Migration, Injection, Staining, Expressing

Journal: Frontiers in Pharmacology

Article Title: Profilin 1 Induces Tumor Metastasis by Promoting Microvesicle Secretion Through the ROCK 1/p-MLC Pathway in Non-Small Cell Lung Cancer

doi: 10.3389/fphar.2022.890891

Figure Lengend Snippet: Schematic diagram of the role of PFN1 in NSCLC metastasis. In the initiation stage of NSCLC, cells with upregulated PFN1 secret more MVs through PFN1 interactions with the ROCK/p-MLC pathway. These MVs contain numerous oncogenenic moleculars, which could enhance migration abilities of PFN1 normal expressed NSCLC cells, and untimately promote progression and metastasis of NSCLC.

Article Snippet: PFN1 siRNA and control scramble siRNA were synthesized by Guangzhou RiboBio Co. (Guangzhou, China), and the sequences are listed in .

Techniques: Migration

Journal: Molecular Medicine

Article Title: Modulation of anti-cardiac fibrosis immune responses by changing M2 macrophages into M1 macrophages

doi: 10.1186/s10020-024-00858-z

Figure Lengend Snippet: NaW increases lysosomal pH-mediated Ca 2+ release and thus enhances glucose uptake. A – G IL-4-conditioned bone marrow-derived macrophages (BMDMs) were treated with or without NaW (2 mM) for 12 h. (A) Scl2a1 expression was measured by real-time PCR. B The mean fluorescence intensity (MFI) level of 2NBDG was measured by flow cytometry. C LysoSensor Green-labeled acidic lysosomes were observed under a fluorescence microscope. Scale bar, 20 μm. D The MFI of the LysoSensor probe was measured by flow cytometry. E , F TMEM175 expression was measured by real-time PCR ( E ) and western blotting ( F ). G – I IL-4-conditioned BMDMs were transfected with TMEM175 siRNAs for 12 h, the MFI of the LysoSensor probe was detected by flow cytometry ( G ), and Nos2 expression was determined by real-time PCR ( H ) and western blotting ( I ). J After Tmem175 overexpression in macrophages, the MFI emitted by 2NBDG was measured by flow cytometry. K IL-4-conditioned BMDMs were treated with or without NaW (2 mM) for 12 h, and cytosolic calcium release was measured by flow cytometry. L , M IL-4-conditioned BMDMs were treated with NH 4 Cl for 5 min, the pH value of lysosomes was measured with a microplate reader ( L ), and the cytosolic calcium release rate was measured by flow cytometry ( M ). N The expression of Mcoln1 and Mcoln2 was analyzed by real-time PCR. O The same as presented in ( A ), except cells were pretreated with 10 nM CsA for 1 h. P , Q The same as presented in ( A ), except the expression and location of TFEB were analyzed by real-time PCR ( P ) and immunofluorescence ( Q ) (scale bar, 20 μm). R , S IL-4-conditioned BMDMs were transfected with Tfeb siRNA, Scl2a1 expression was measured by real-time PCR, and the MFI of 2NBDG was measured by flow cytometry. Unless otherwise specified, n = 3 biologically independent experiments. The data are presented as the mean ± SEM. P values were calculated using one-way ANOVA. *P < 0.05; **P < 0.01; ***P < 0.001

Article Snippet: Short interfering RNAs (siRNAs) targeting mouse Pygl , Tmem175 , and Tfeb and negative control siRNAs (NC) were purchased from RiboBio (Guangzhou, China). siRNA (50 nM) was transfected into macrophages using LipofectamineTM RNAiMAX Transfection Reagent according to the manufacturer’s instructions.

Techniques: Derivative Assay, Expressing, Real-time Polymerase Chain Reaction, Fluorescence, Flow Cytometry, Labeling, Microscopy, Western Blot, Transfection, Over Expression, Immunofluorescence

Journal: Frontiers in Genetics

Article Title: Elevated Serum FGG Levels Prognosticate and Promote the Disease Progression in Prostate Cancer

doi: 10.3389/fgene.2021.651647

Figure Lengend Snippet: FGG knockdown inhibits PCa cell proliferation in vivo . (A) The efficiency of the shRNA. (B) Bioluminescence images of the resultant DU145 tumors in the NC and FGG-KD groups. (C) Subcutaneous tumors were excised and weighed 4 weeks after inoculation. (D) Relative tumor volumes of the NC and FGG-KD groups. (E) Representative IHC and HE staining images of the NC and FGG-KD subcutaneous tumors. * p < 0.05, ** p < 0.01.

Article Snippet: Cells were seeded in six-well plates and were incubated with antibiotic-free medium half an hour before transfection. si-FGG, si-STAT3, and non-targeting control siRNA (si-control) (Synbio Technologies, China) were transfected in Opti-MEM medium (Invitrogen) using X-tremeGENE TM siRNA Transfection Reagent (Roche, Germany) for 6 h according to the manufacturer’s instructions.

Techniques: Knockdown, In Vivo, shRNA, Staining

Journal: BMC Cancer

Article Title: MICAL1 controls cell invasive phenotype via regulating oxidative stress in breast cancer cells

doi: 10.1186/s12885-016-2553-1

Figure Lengend Snippet: Active form of RAB35 binds to MICAL1. a Representative micrographs of MDA-MB-231 and MCF-7 cells stained for RAB35 (green) and MICAL1 expression (red) by immunofluorescence assay. Scale bar, 10 μm. b Coimmunoprecipitation experiments were performed with HEK293T cells cotransfected with HA-tagged MICAL1 and GFP-tagged RAB35. c MCF-7 cells were transfected with GFP-tagged RAB35 (WT), RAB35 (DN) or RAB35 (CA), and then immunoprecipitated with anti-GFP antibody, followed by immunoblotting analysis for RAB35 and MICAL1. d Binding of endogenous RAB35 to MICAL1 was detected in MDA-MB-231 cells and MCF-7 cells by coimmunoprecipitation experiments. n = 3 for all experiments

Article Snippet: The sequences of small interfering RNA (siRNA) for MICAL1 were as follows: #1, 5′-GUCUCUGCCUUUGACUUCATT-3′, #2, 5′-CUGCAGAACAUUGUGUACUTT-3′, and #3, 5′-CUCGGUGCUAAGAAGUUCUTT-3′; siRNA for RAB35 was: 5′-GCAGCAACAACAGAACGAUTT-3′ and the sequence of control siRNA was 5′-UUCUCCGAACGUGUCACGUTT-3′ (GenePharma, Shanghai, China).

Techniques: Staining, Expressing, Immunofluorescence, Transfection, Immunoprecipitation, Western Blot, Binding Assay

Journal: BMC Cancer

Article Title: MICAL1 controls cell invasive phenotype via regulating oxidative stress in breast cancer cells

doi: 10.1186/s12885-016-2553-1

Figure Lengend Snippet: Effect of RAB35 on breast cancer cell invasion. a RAB35 and MICAL1 expressions were examined by immunoblotting in several types of malignant breast cancer cell line. b MDA-MB-231 cells were incubated with EGF (10 ng/mL) for indicated times, and analyzed for RAB35 activity by pulldown assays. * P < 0.05 in the cultures with EGF relative to the cultures without EGF. c MCF-7 cells were transfected with negative control siRNA or siRAB35. 48 h later, total protein extracts from cells were analyzed by immunoblotting analysis for RAB35 expression. Western blot bands corresponding to RAB35 were quantified and normalized against GAPDH level. **: P < 0.01 in the siRAB35 cells relative to control siRNA cells. d MCF-7 cells transfected with control siRNA or siRAB35, and the quantification of cell invasion rate was performed. *: P < 0.05 in the siMICAL1 cells relative to control siRNA cells. e Invasion assays results showed that knockdown of MICAL1 delayed cell invasion in RAB35 (CA)-expressing MDA-MB-231 cells. *: P < 0.05. **: P < 0.01

Article Snippet: The sequences of small interfering RNA (siRNA) for MICAL1 were as follows: #1, 5′-GUCUCUGCCUUUGACUUCATT-3′, #2, 5′-CUGCAGAACAUUGUGUACUTT-3′, and #3, 5′-CUCGGUGCUAAGAAGUUCUTT-3′; siRNA for RAB35 was: 5′-GCAGCAACAACAGAACGAUTT-3′ and the sequence of control siRNA was 5′-UUCUCCGAACGUGUCACGUTT-3′ (GenePharma, Shanghai, China).

Techniques: Western Blot, Incubation, Activity Assay, Transfection, Negative Control, Expressing, Control, Knockdown

Journal: BMC Cancer

Article Title: MICAL1 controls cell invasive phenotype via regulating oxidative stress in breast cancer cells

doi: 10.1186/s12885-016-2553-1

Figure Lengend Snippet: RAB35/MICAL1 regulates ROS generation. a Effects of RAB35 and MICAL1 on H 2 O 2 generation. MCF-7 cells transfected with siRAB35 or siMICAL1 were in serum-free media overnight. Representative micrographs of those cells incubated with EGF (10 ng/mL for 30 min) and stained with CM 2 -DCFHDA. *: P < 0.05 in the cultures with EGF relative to the cultures without EGF. # : P < 0.05 in the cultures with EGF plus siRAB35 or siMICAL1 relative to the cultures with EGF. Scale bar, 100 μm. b Effects of RAB35 and MICAL1 on O 2 − generation in MCF-7 cells. *: P < 0.05 in the cultures transfected with siRAB35 or siMICAL1 relative to the cultures with control siRNA. c Quantification of O 2 − levels in the cells transfected with HA–MICAL1, HA tagged MICAL1 FAD-LIM domain and CC domain. *: P < 0.05 in the cells transfected with HA–MICAL1 or HA–FAD-LIM relative to cells transfected with vector. # : P < 0.05 in the cells transfected with CC domain relative to cells transfected with HA–MICAL1 or HA–FAD-LIM domain

Article Snippet: The sequences of small interfering RNA (siRNA) for MICAL1 were as follows: #1, 5′-GUCUCUGCCUUUGACUUCATT-3′, #2, 5′-CUGCAGAACAUUGUGUACUTT-3′, and #3, 5′-CUCGGUGCUAAGAAGUUCUTT-3′; siRNA for RAB35 was: 5′-GCAGCAACAACAGAACGAUTT-3′ and the sequence of control siRNA was 5′-UUCUCCGAACGUGUCACGUTT-3′ (GenePharma, Shanghai, China).

Techniques: Transfection, Incubation, Staining, Control, Plasmid Preparation

Journal: BMC Cancer

Article Title: MICAL1 controls cell invasive phenotype via regulating oxidative stress in breast cancer cells

doi: 10.1186/s12885-016-2553-1

Figure Lengend Snippet: Effects of RAB35 and MICAL1 on ROS-modulated Akt activity. a MCF-7 cells transfected with siRAB35 or siMICAL1, and protein levels of P-Akt and Akt were examined. Western blot bands corresponding to P-Akt were quantified and normalized against Akt levels. *: P < 0.05 in the cultures transfected with siRAB35 or siMICAL1 relative to the cultures with control siRNA. b MCF-7 cells were transfected with MICAL1 or RAB35 (CA) plasmids, and the total cellular proteins were extracted and analyzed for expressions of P-Akt and Akt by immunoblotting assays. *: P < 0.05 in the cells transfected with HA–MICAL1 or GFP–RAB35 (CA) relative to cells transfected with the corresponding vector. c MCF-7 cells were treated with 2 mM NAC for 1 h, and then the total protein extracts from cells were analyzed by immunoblotting assays for P-Akt and Akt expression. d Effect of NAC on cell invasion. After cultured in serum-free medium overnight, MCF-7 cells were pretreated with 2 mM NAC for 1 h, and then were stimulated with EGF (10 ng/mL) or 10 % FBS for 24 h. Quantifications of cells on the lower surface of the membrane were performed and shown. *: P < 0.05, **: P < 0.01 in the cultures with NAC relative to the cultures without NAC. e Invasion assays results showed that LY294002 and NAC delayed cell invasion in MICAL1-expressing MCF-7 cells. *: P < 0.05. **: P < 0.01

Article Snippet: The sequences of small interfering RNA (siRNA) for MICAL1 were as follows: #1, 5′-GUCUCUGCCUUUGACUUCATT-3′, #2, 5′-CUGCAGAACAUUGUGUACUTT-3′, and #3, 5′-CUCGGUGCUAAGAAGUUCUTT-3′; siRNA for RAB35 was: 5′-GCAGCAACAACAGAACGAUTT-3′ and the sequence of control siRNA was 5′-UUCUCCGAACGUGUCACGUTT-3′ (GenePharma, Shanghai, China).

Techniques: Activity Assay, Transfection, Western Blot, Control, Plasmid Preparation, Expressing, Cell Culture, Membrane

Journal: American Journal of Cancer Research

Article Title: HDAC6/FOXP3/HNF4α axis promotes bile acids induced gastric intestinal metaplasia

doi:

Figure Lengend Snippet: Sequence of primers used in the study

Article Snippet: The small interference RNAs (siRNAs) to HDAC6, FOXP3 and HNF4α were purchased from Genepharma (China).

Techniques: Sequencing

Journal: American Journal of Cancer Research

Article Title: HDAC6/FOXP3/HNF4α axis promotes bile acids induced gastric intestinal metaplasia

doi:

Figure Lengend Snippet: Sequence of primers used in the study

Article Snippet: The small interference RNAs (siRNAs) to HDAC6, FOXP3 and HNF4α were purchased from Genepharma (China).

Techniques: Sequencing

Journal: American Journal of Cancer Research

Article Title: HDAC6/FOXP3/HNF4α axis promotes bile acids induced gastric intestinal metaplasia

doi:

Figure Lengend Snippet: FOXP3 was decreased in gastric intestinal metaplasia (IM). A. qRT-PCR results of expression of FOXP3 in 10 paired IM and surrounding tissues. B. Comparison of FOXP3, HDAC6 and HNF4α mRNA expression in the biopsies. **P < 0.01.

Article Snippet: The small interference RNAs (siRNAs) to HDAC6, FOXP3 and HNF4α were purchased from Genepharma (China).

Techniques: Quantitative RT-PCR, Expressing, Comparison

Journal: American Journal of Cancer Research

Article Title: HDAC6/FOXP3/HNF4α axis promotes bile acids induced gastric intestinal metaplasia

doi:

Figure Lengend Snippet: HDAC6 induces extensive chromatin modifications at FOXP3 loci. A. Histone acetyltransferase (HATs) and histone deacetylases (HDACs) affect the dynamics of chromatin remodeling during gene transcription. B. Schematic diagram depicting FOXP3 locus structure and the sequences in the FOXP3 promoter and CNS regions. C. ChIP assays for H3Ac, H3K9Ac, H3K27Ac, H4Ac modifications in FOXP3 locus. IgG serves as immunoprecipitation control. Student’s t-test. D. ChIP assays for HDAC6 in FOXP3 promoter and CNS regions in GES-1 cells. Student’s t-test. E. FOXP3 and HNF4α protein levels was examined by western blot analysis in AGS cells treated with EP300-expressing vectors or control. *P < 0.05, **P < 0.01. N.S., not significant.

Article Snippet: The small interference RNAs (siRNAs) to HDAC6, FOXP3 and HNF4α were purchased from Genepharma (China).

Techniques: Immunoprecipitation, Control, Western Blot, Expressing

Journal: American Journal of Cancer Research

Article Title: HDAC6/FOXP3/HNF4α axis promotes bile acids induced gastric intestinal metaplasia

doi:

Figure Lengend Snippet: FOXP3 negatively regulated HNF4α transcription. A. FOXP3 inversely regulated HNF4α at both mRNA and protein levels. Student’s t-test. B. The promoter activity of HNF4α was enhanced by DCA and siFOXP3 treatment in GES-1 cells. Student’s t-test. C. A ChIP assay demonstrated the direct binding of FOXP3 to the HNF4α promoter in GES-1 cells. Student’s t-test. D. AGS cells were co-infected with FOXP3-expressing vectors and HNF4α-expressing vectors. The expression of intestinal markers was detected by immunoblot. E. Upregulated intestinal markers expression induced by siFOXP3 was attenuated by siHNF4α in GES-1 cells. F. A schematic model of miR-1/HDAC6/FOXP3/HNF4α pathway in gastric cells. In response to BA, miR-1 inhibition promotes HNF4α and HDAC6, which increase the transcription of intestinal markers including MUC2, KLF4 and CDX2. **P < 0.01. N.S., not significant.

Article Snippet: The small interference RNAs (siRNAs) to HDAC6, FOXP3 and HNF4α were purchased from Genepharma (China).

Techniques: Activity Assay, Binding Assay, Infection, Expressing, Western Blot, Inhibition

Journal: American Journal of Cancer Research

Article Title: HDAC6/FOXP3/HNF4α axis promotes bile acids induced gastric intestinal metaplasia

doi:

Figure Lengend Snippet: HDAC6/FOXP3/HNF4α loop in mice. A. Experimental strategy of establishing gastric intestinal metaplasia model in mice. Active Hnf4α expression was induced after tamoxifen treatment. B. Representative pictures of IHC staining for FOXP3, HDAC6 and HNF4α at the SCJ of WT and Rosa26Hnf4α mice with or without DCA treatment. Scale bars: 20 µm (bottom). C. Expression of these molecules in the gastric antrum of four groups of mice. Scale bars: 20 µm (bottom).

Article Snippet: The small interference RNAs (siRNAs) to HDAC6, FOXP3 and HNF4α were purchased from Genepharma (China).

Techniques: Expressing, Immunohistochemistry

Journal: American Journal of Cancer Research

Article Title: HDAC6/FOXP3/HNF4α axis promotes bile acids induced gastric intestinal metaplasia

doi:

Figure Lengend Snippet: Histopathological changes of gastric mucosa in mice (χ 2 test)

Article Snippet: The small interference RNAs (siRNAs) to HDAC6, FOXP3 and HNF4α were purchased from Genepharma (China).

Techniques: