Journal: The American Journal of Pathology
Article Title: Smooth Muscle α-Actin Deficiency Leads to Decreased Liver Fibrosis via Impaired Cytoskeletal Signaling in Hepatic Stellate Cells
doi: 10.1016/j.ajpath.2019.07.019
Figure Lengend Snippet: Cellular features and COL1 expression of smooth muscle (SM) α-actin–deficient hepatic stellate cells (HSCs). A: HSCs from Acta2+/+ and Acta2−/− mice were cultured in standard medium as indicated, and images were captured using darkfield phase microscopy. The arrows indicate an individual HSC, and the insets show a higher magnification (representative images are shown from >10 others). B: Cells were cultured as in A for 3 days and subjected to Oil Red O staining. The arrows indicate cytoplasmic lipid droplets in HSCs (representative images are shown of >10 others). C: Cells were as in A, and total RNA was isolated. Real-time quantitative PCR was performed to detect the indicated mRNAs. D: Cells were cultured as in A for 5 days and then incubated in 0.5% serum medium with or without 10 ng/mL TGF-β or 20 μmol/L ET-1 for 24 hours. Total RNA was isolated, and COL1α1 mRNA levels were measured by real-time quantitative PCR. E: Rat hepatic stellate cells were exposed to Ad-control or Ad-SM α-actin [25 or 50 multiplicity of infection (MOI)] for 3 days in 0.5% serum medium and then incubated in 0.5% serum medium for an additional 1 day; whole-cell lysates were subjected to immunoblotting as indicated, and the quantitative data were presented graphically. n = 3. *P < 0.05, **P < 0.01, and ***P < 0.001 for Acta2+/+ versus Acta2−/−. Scale bars: 20 μm (A); 40 μm (B). Original magnification, ×20 (insets).
Article Snippet: Anti-COL1 antibody (600-401-103-0.5) was from Rockland (Gilbertsville, PA).
Techniques: Expressing, Cell Culture, Microscopy, Staining, Isolation, Real-time Polymerase Chain Reaction, Incubation, Infection, Western Blot
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![Cellular features and <t>COL1</t> expression of smooth muscle (SM) α-actin–deficient hepatic stellate cells (HSCs). A: HSCs from Acta2+/+ and Acta2−/− mice were cultured in standard medium as indicated, and images were captured using darkfield phase microscopy. The arrows indicate an individual HSC, and the insets show a higher magnification (representative images are shown from >10 others). B: Cells were cultured as in A for 3 days and subjected to Oil Red O staining. The arrows indicate cytoplasmic lipid droplets in HSCs (representative images are shown of >10 others). C: Cells were as in A, and total RNA was isolated. Real-time quantitative PCR was performed to detect the indicated mRNAs. D: Cells were cultured as in A for 5 days and then incubated in 0.5% serum medium with or without 10 ng/mL TGF-β or 20 μmol/L ET-1 for 24 hours. Total RNA was isolated, and COL1α1 mRNA levels were measured by real-time quantitative PCR. E: Rat hepatic stellate cells were exposed to Ad-control or Ad-SM α-actin [25 or 50 multiplicity of infection (MOI)] for 3 days in 0.5% serum medium and then incubated in 0.5% serum medium for an additional 1 day; whole-cell lysates were subjected to immunoblotting as indicated, and the quantitative data were presented graphically. n = 3. *P < 0.05, **P < 0.01, and ***P < 0.001 for Acta2+/+ versus Acta2−/−. Scale bars: 20 μm (A); 40 μm (B). Original magnification, ×20 (insets).](https://pub-med-central-images-cdn.bioz.com/pub_med_central_ids_ending_with_2116/pmc06902116/pmc06902116__gr3.jpg)
