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:

Journal: Frontiers in Pharmacology

Article Title: Nonmuscle Myosin Heavy Chain ⅡA-Mediated Exosome Release via Regulation of the Rho-Associated Kinase 1/Myosin Light Chains/Actin Pathway

doi: 10.3389/fphar.2020.598592

Figure Lengend Snippet: Detection of NMMHC ⅡA, NMMHC ⅡB, and NMMHC ⅡC in exosomes extracted from microglial cells. For transfection, microglial cells were sub-cultured in 24-well plates at a density of 3 × 10 5 cells/ml. Cells were transfected with siRNA-MYH9, siRNA-MYH10, or siRNA-MYH14 at a final concentration of 100 nM for 48 h. The extracted exosomes were grouped based on whether they were extracted from control microglial cells, microglial cells transfected with siRNA, microglial cells transfected with siRNA negative control, microglial cells stimulated by 1 mg/ml LPS for 24 h, microglial cells transfected with siRNA followed by 24 h LPS stimulation, or microglial cells transfected with siRNA negative control followed by 24 h LPS stimulation. The NMMHC ⅡA, NMMHC ⅡB, and NMMHC ⅡC protein expression levels for each group of exosomes were examined by western blotting. Western blots were quantified. Nine independent experiments were analyzed. A histogram depicted the quantitative representations of the protein expression levels of CD9 and CD81 for each group. The data were averages with SD, n = 9. ** p < 0.01 vs. control microglial cells; ## p < 0.01 vs. microglial cells stimulated with LPS.

Article Snippet: NMMHC ⅡA siRNA, NMMHC ⅡB siRNA, NMMHC ⅡC siRNA, and control non-specific siRNA were synthesized by the Shanghai Biotechnology Corporation (China). siRNA transfection was performed using an ExFect Transfection Reagent (Shanghai Biotechnology Corporation, China) according to the manufacturer’s instructions.

Techniques: Transfection, Cell Culture, Concentration Assay, Negative Control, Expressing, Western Blot

Journal: Frontiers in Pharmacology

Article Title: Nonmuscle Myosin Heavy Chain ⅡA-Mediated Exosome Release via Regulation of the Rho-Associated Kinase 1/Myosin Light Chains/Actin Pathway

doi: 10.3389/fphar.2020.598592

Figure Lengend Snippet: The NMMHC ⅡA-regulated ROCK1/MLC/actin pathway might be the potential mechanism for exosome release. The exosomes were divided on the basis of whether they were extracted from control microglial cells, microglial cells incubated with 1 mg/ml LPS for 24 h, and microglial cells transfected with siRNA-MYH9 at a final concentration of 100 nM for 48 h. (A) Microglial cells were immunofluorescently stained with anti-NMMHC ⅡA, anti-ROCK1, anti-MLC, or anti-actin, together with CD11b (green) and DAPI for nuclei (blue). Stained cells were examined by confocal microscopy. Scale bars = 10 μm. A histogram depicted the quantitative fluorescence intensity from five vision fields from nine independent experiments for each group of microglial cells. (B) The protein expression levels of anti-NMMHC ⅡA, anti-ROCK1, anti-MLC, and anti-actin were examined by western blotting. Western blots were quantified. A histogram depicted the quantitative representations of the protein expression levels of NMMHC ⅡA, ROCK1, MLC, and actin for each group. Nine independent experiments were analyzed. The data were averages with SD, n = 9. ** p < 0.01 vs. control microglial cells; ## p < 0.01 vs. microglial cells cultured for 24 h.

Article Snippet: NMMHC ⅡA siRNA, NMMHC ⅡB siRNA, NMMHC ⅡC siRNA, and control non-specific siRNA were synthesized by the Shanghai Biotechnology Corporation (China). siRNA transfection was performed using an ExFect Transfection Reagent (Shanghai Biotechnology Corporation, China) according to the manufacturer’s instructions.

Techniques: Incubation, Transfection, Concentration Assay, Staining, Confocal Microscopy, Fluorescence, Expressing, Western Blot, Cell Culture