Journal: PLoS ONE
Article Title: The Hepatic Raldh1 Expression Is elevated in Zucker Fatty Rats and Its Over-Expression Introduced the Retinal-Induced Srebp-1c Expression in INS-1 Cells
Figure Lengend Snippet: The over-expression of RALDH1 resulted in the RAL-mediated induction of Srebp-1c in 833/15 INS-1 cells. A. The adenovirus-mediated Raldh1 mRNA expression. B. Immuno-blot of the over-expression of RALDH1 protein in INS-1 cells. Whole cell lysates (50 µg/sample) of the control cells (lane 1), cells infected by the indicated pfu of Ad-β-gal (lane 2) or Ad-Raldh1 (lanes 3–5) were separated in 8% SDS protein gels, and transferred to the PVDF membranes. Primary antibodies to RALDH1 (1∶1000 dilution in TBST containing 5% dry milk), and to β-Actin (1∶1000 dilution in TBST containing 5% bovine serum albumin) were recognized by goat anti-rabbit IgG conjugated to horseradish peroxidase, and visualized by chemiluminescence. The films were scanned and presented as described in the Material and Methods . C. RAL only induced Srebp-1c expression in cells over-expressing RALDH1, but not β-gal. D. RA induced Srebp-1c expression in cells over-expressing either β-gal or RALDH1. Results were presented as means ± SD of fold inductions (* for comparing the different dosages of RAL in cells infected by Ad-Raldh1 using one way ANOVA, n = 3, all p
Article Snippet: Proteins (40 μg/lane) in whole cell lysates were separated on SDS-PAGE, transferred to BIO-RAD Immuno-Blot PVDF membrane (Hercules, CA) and detected with primary antibodies to RALDH1 (for , catalog #2052-1, Epitomics, Burlingame, CA 94010), RALDH1 (for , catalog #AP1465a, Abgent, San Diego, CA 92121), CYP26A1 (catalog # CYP26A11-A, Alpha Diagnostics International, TX 78244), and β-Actin (#4970 s, Cell Signaling Technology, Danvers, MA 01923) according to the protocols provided by the manufacturers.
Techniques: Over Expression, Expressing, Infection