Journal: Cell Death & Disease

Article Title: MiR-130a-3p attenuates activation and induces apoptosis of hepatic stellate cells in nonalcoholic fibrosing steatohepatitis by directly targeting TGFBR1 and TGFBR2

doi: 10.1038/cddis.2017.10

Figure Lengend Snippet: miR-130a-3p inhibited cell proliferation and growth, and induced apoptosis in activated HSCs. (a) The miR-130a-3p mimics regulated the expression of miR-130a-3p. qRT-PCR validated the expression of miR-130a-3p after transfection with the miR-130a-3p mimics. The miR-130a-3p mimics significantly upregulated the expression of miR-130a-3p. Values represent the mean±S.D., *** P <0.001 compared with control. (b) miR-130a-3p expression was examined by real-time qRT-PCR at 0, 24, 48, 72 h after transfection with the miR-130a-3p mimics. Values represent the mean±S.D. (** P <0.01). ( c ) miR-130a-3p inhibited HSC-T6 cell growth as determined by an MTT assay. Values represent the mean±S.D., ** P <0.01, *** P <0.001 compared with control. ( d ) Annexin V/PI combined with flow cytometry. ( e ) The mRNA expression of caspase-3, caspase-9, and PARP1 and ( f ) the protein expression of cleaved caspase-3, caspase-9, and PARP1 were increased after transfection with the miR-130a-3p mimics. β -actin was used as a loading control. Values represent the mean±S.D., *** P <0.001 compared with control

Article Snippet: After being blocked for 60 min with buffer containing 0.1% Tween-20 and 5% milk, the membranes were incubated overnight at 4 °C with primary antibodies against α -SMA, Col-1, Col-4 (Bioss, Beijing, China), Smad2, Smad3 (Novus Biologicals, Novus Biologicals, Littleton, CO, USA), TGFBR1 (Abcam), TGFBR2 (Novus Biologicals), MMP-2, MMP-9 (ProteinTech Group, Chicago, IL, USA), TGF- β 1 , caspase-3, caspase-9, and PARP1 (Novus Biologicals, Littleton, CO, USA).

Techniques: Expressing, Quantitative RT-PCR, Transfection, Control, MTT Assay, Flow Cytometry

Journal: Cell Death & Disease

Article Title: MiR-130a-3p attenuates activation and induces apoptosis of hepatic stellate cells in nonalcoholic fibrosing steatohepatitis by directly targeting TGFBR1 and TGFBR2

doi: 10.1038/cddis.2017.10

Figure Lengend Snippet: Primers for quantitative real-time PCR analysis

Article Snippet: After being blocked for 60 min with buffer containing 0.1% Tween-20 and 5% milk, the membranes were incubated overnight at 4 °C with primary antibodies against α -SMA, Col-1, Col-4 (Bioss, Beijing, China), Smad2, Smad3 (Novus Biologicals, Novus Biologicals, Littleton, CO, USA), TGFBR1 (Abcam), TGFBR2 (Novus Biologicals), MMP-2, MMP-9 (ProteinTech Group, Chicago, IL, USA), TGF- β 1 , caspase-3, caspase-9, and PARP1 (Novus Biologicals, Littleton, CO, USA).

Techniques: Real-time Polymerase Chain Reaction