Journal: bioRxiv

Article Title: Microsecond pulse electrical stimulation modulates cell migration

doi: 10.1101/2022.10.23.513372

Figure Lengend Snippet: (a) A graphical illustration showed the effects of μsPEF (pulse width:20 μs, frequency: 10 Hz, duration: 5 s) on the skin wound, promoting cell migration and extracellular matrix remodeling. (b) Representative time-lapse images showing the fibroblasts morphology after μsPEF (i.e., 750 and 1500 V/cm) treatment of different intensity at different time points (i.e., 0, 1 and 2 h). Detached cells are marked by yellow arrow. Scale bar, 50 μm. (c) The line graph representing the cell migration average speed per 1 h from 0 to 12 h after different intensity μsPEF (i.e., 750 and 1500 V/cm) treatment (blue circle: control; orange square: 750 V/cm; pink triangle: 1500 V/cm). Results are presented as mean ± standard deviation with 95% CI (n CTRL =42 cells, n 750 v/cm =48 cells, n 1500 v/cm =47 cells); * p<0.05, ** p<0.01, *** p<0.001, **** p<0.0001 versus control by one-way ANOVA for multiple comparisons. (d) Box plot showing the average cell migration speed with different intensity μsPEF (i.e., 750 and 1500 V/cm) and control treatments in 24 h. Results are presented as mean ± standard deviation with 95% CI (n CTRL =470 cells, n 750 V/cm =979 cells, n 1500 V/cm =535 cells). (e) Effects of μsPEF on the secretion of COLA2. The level of collagen type I α2 in cellular supernatants was measured after 48 h. The concentration of COLA2 was 0.109 ± 0.018 ng/mL in the control group, 0.257 ± 0.058 ng/mL in the 750 V/cm group and 0.363 ± 0.034 ng/mL in the 1500 V/cm group. Results are presented as mean ± standard deviation with 95% CI (n=5); * p<0.05, ** p<0.01, *** p<0.001, **** p<0.0001 versus control by one-way ANOVA for multiple comparisons. (f) Effects of μsPEF on the expression of FGF2. The level of FGF2 in cellular supernatants was measured after 48 h. The concentration of FGF2 was 44.139 ± 0.360 ng/mL in the control group, 48.012 ± 1.488 ng/mL in the 750 V/cm group and 48.523 ± 1.944 ng/mL in the 1500 V/cm group. Results are presented as mean ± standard deviation with 95% CI (n=3); ns=0.7329, * p<0.05, ** p<0.01, *** p<0.001, **** p<0.0001 versus control by one-way ANOVA for multiple comparisons.

Article Snippet: Then, the cells were cultured in serum-free medium for 48 h. The content of type I α collagen and basic fibroblast growth factor (FGF-2) in the supernatant were measured using commercially available Human COL1A2 (Collagen Type I Alpha 2) ELISA Kit (Elabscience, Wuhan, China) and Human bFGF/FGF2 (Basic Fibroblast Growth Factor) ELISA Kit (Elabscience, Wuhan, China) according to the manufacturer’s protocol respectively.

Techniques: Migration, Control, Standard Deviation, Concentration Assay, Expressing

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

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: A proposed molecular mechanism underlying the effect of smooth muscle (SM) α-actin on hepatic myofibroblast differentiation and liver fibrogenesis. SM α-actin is encoded by Acta2 and transactivated by serum response factor (SRF) during hepatic stellate cell activation. This leads to actin polymerization and stress fiber formation, which not only serves as the functional actin cytoskeleton, but also plays an important role in transmission of mechanical signals to the nucleus to regulate COL1 expression. Additionally, signaling mediated by important soluble factors such as transforming growth factor-β (TGF-β) and ET-1 is impaired in Acta2-deficient hepatic stellate cells, contributing to reduced expression of COL1 and liver fibrosis in Acta2-deficient mice.

Article Snippet: Anti-COL1 antibody (600-401-103-0.5) was from Rockland (Gilbertsville, PA).

Techniques: Activation Assay, Functional Assay, Transmission Assay, Expressing