Journal: Bioengineered

Article Title: Astragalus polysaccharide improve the proliferation and insulin secretion of mouse pancreatic β cells induced by high glucose and palmitic acid partially through promoting miR-136-5p and miR-149-5p expression

doi: 10.1080/21655979.2021.1996314

Figure Lengend Snippet: Silencing miR-136-5p and miR-149-5p gene reduced cell survival, insulin secretion, and PDX1 mRNA expression and promoted cell apoptosis in MIN6 cells co-treated with HG+PA and 200 μg/ml astragalus polysaccharide (APS). (a) Expression of miR-136-5p and miR-149-5p was measured by RT-qPCR after transfection at 24 h. (b) The proliferation was measured by CCK8 after transfection at 24, 48, 72 h, then the survival rate was calculated according to the OD value of NC inhibitor group. (c) PDX1 mRNA expression was measured by RT-qPCR after transfection at 48 h. (d) Insulin secretion was measured by ELISA after transfection at 48 h. (e-f) Apoptosis was measured by flow cytometry after transfection at 48 h. # P < 0.05, vs NC inhibitor group

Article Snippet: The negative control inhibitor (NC inhibitor group), mmu-miR-136-5p, and mmu-miR-149-5p inhibitors (miR-136-5p inhibitor and miR-149-5p inhibitor groups) were purchased from Shanghai GenePharma (Shanghai, China).

Techniques: Expressing, Quantitative RT-PCR, Transfection, Enzyme-linked Immunosorbent Assay, Flow Cytometry

Journal: Bioengineered

Article Title: Astragalus polysaccharide improve the proliferation and insulin secretion of mouse pancreatic β cells induced by high glucose and palmitic acid partially through promoting miR-136-5p and miR-149-5p expression

doi: 10.1080/21655979.2021.1996314

Figure Lengend Snippet: EFHD2 is the potential target genes of miR-136-5p and miR-149-5p. (a) The potential target genes of miR-136-5p and miR-149-5p were predicted by Starbase 3.0. And different-expressed mRNAs in β cell between non-diabetic condition and diabetic condition were analyzed by GEO2R in the GSE20966 dataset of Gene Expression Omnibus. Intersection of 3 sets of results was shown. (b) The heat map about the different-expressed mRNAs in β cell between non-diabetic condition and diabetic condition which analyzed by GEO2R in the GSE20966 dataset. Red: overexpression; Green: significant downexpression. (c) The EFHD2 expression was analyzed in GSE20966 dataset. (d) the bind site between miR-136-5p, miR-149-5p and 3′-UTR. And this bind was verified by luciferase reporter assay. The renilla/firefly ratio was significantly lower in miR-136-5p/miR-149-5p mimic + WT EFHD2 group than that in NC mimic + WT EFHD2 group, whereas the renilla/firefly ratio had no significantly changed between NC mimic + Mut EFHD2 group and miR-136-5p/miR-149-5p mimic + Mut EFHD2 group, suggesting that miR-136-5p and miR-149-5p can bind with the 3′-UTR of EFHD2. (e) EFHD2 expression was measured by western blot

Article Snippet: The negative control inhibitor (NC inhibitor group), mmu-miR-136-5p, and mmu-miR-149-5p inhibitors (miR-136-5p inhibitor and miR-149-5p inhibitor groups) were purchased from Shanghai GenePharma (Shanghai, China).

Techniques: Expressing, Over Expression, Luciferase, Reporter Assay, Western Blot

Journal: Oncotarget

Article Title: TRIM11, a direct target of miR-24-3p, promotes cell proliferation and inhibits apoptosis in colon cancer

doi: 10.18632/oncotarget.13550

Figure Lengend Snippet: A . Western blot analysis of TRIM11 protein levels after transfection of miRNAs mimics in HCT116 cells. B . Luciferase activity was measured 24 h after transfection of miRNAs mimics in 293T cells. Renilla luciferase was used for normalization. The bars correspond to the mean± standard error, and the p-value was calculated using Student's t-test. *P<0.05. C . The sequence of miR-24-3p and the 7-mer binding site in 3’ UTR of TRIM11 mRNA. Red letters are the mutated nucleotides in the seed sequence of 3’UTR. D . Mutant luciferase activity was measured 24 h after transfection of miR-24-3p mimics in 293T cells. E, F . The levels of miR-24-3p and TRIM11 were detected after transfection of miR24-3p mimics in HCT116 cells. G . Western blot analysis of TRIM11 protein levels after transfection of miR24-3p mimics. H . Western blot analysis of TRIM11 protein levels after transfection of miR24-3p inhibitors.

Article Snippet: The miRNA mimics (miR-24-3p, miR-101-3p, miR-148a, miR-15a, miR-504, miR-329, miR-370, miR-149, miR-320a, miR-193b, miR-544a, miR-150, miR-124-3p), miRNA control, and miRNA inhibitors for miR-24-3p were purchased from Ribobio (Ribobio, Guangzhou, China).

Techniques: Western Blot, Transfection, Luciferase, Activity Assay, Sequencing, Binding Assay, Mutagenesis