sc 5310  (Santa Cruz Biotechnology)

Journal: Molecular Metabolism

Article Title: α-Parvin promotes glucose uptake and metabolism in skeletal muscle with minimal influence on hepatic insulin sensitivity

doi: 10.1016/j.molmet.2026.102322

Figure Lengend Snippet: α-Parvin regulates skeletal muscle actin remodeling and GLUT4 membrane recruitment. A ) RhoA and Rac1 activity assays were determined in the basal and insulin-stimulated state in gastrocnemius muscle. B) Immunoblotting for total and phosphorylated cofilin was determined in gastrocnemius muscle in basal and insulin-stimulated conditions. The phosphorylated cofilin densitometry was normalized to total cofilin. C ) Sections of gastrocnemius muscle were collected from the mice indicated and fixed in a solution of 10% formalin paraformaldehyde in phosphate buffered saline or snap frozen in isopentane at liquid nitrogen temperature and affixed to cork blocks with OCT. Fixed tissues were embedded in paraffin and sectioned and frozen tissues were cryo-sectioned and mounted. Antibodies against Cav3, GLUT4, and AF-647–phalloidin (F-actin) were used to determine GLUT4 cellular localization. Confocal muscle images were collected with confocal microscopy at super-resolution using a Zeiss LSM 980 confocal microscope equipped with an inverted Axio Observer 7 and Airyscan 2 detector. D ) Airyscan super-resolution images were acquired under identical settings for all groups and images. Acquisition and 2D Airyscan processing of acquired images was done using ZEN Blue software (Carl Zeiss). Line Scan profiles of fluorescence intensity along an annotated line was performed using the plot profile function in Fiji/ImageJ. E ) Gastrocnemius muscle tissue was fractionated to separate cytosolic and plasma membrane fractions for immunoblotting. Purity of fractions were determined by the plasma membrane protein Na/K Atpase. N = 4 to 6 mice per group. Panel A and B, Two-way ANOVA with genotype and insulin as factors was run to test for between group differences. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Antibodies against Cav3 (Santa Cruz #sc55518), GLUT4 (Abcam #ab33780), α-Parvin (Cell Signaling (CST) #4026), HKII (Abcam # ab227198), VDAC1 (MilliporeSigma #SAB5201374) were used and F-actin was visualized using AF-647–phalloidin (Thermo, #A22287).

Techniques: Membrane, Activity Assay, Western Blot, Saline, Confocal Microscopy, Microscopy, Software, Fluorescence, Clinical Proteomics