Fig. 5

Western blot analyses of perilipin 1 or 2 suppressed RINm5F and INS-1E cells. Perilipin 1 and 2 expression in RINm5F (a, c) and INS-1E (b, d) cells was stably suppressed by the shRNA technique after lentiviral transduction. Cells were incubated with 100 μM palmitic acid (PA), 100 μM oleic acid (OA), a combination of both, or 100 μM gondoic acid (GA) or without NEFAs (Con) for 24 h. Thereafter, 30 μg protein of total whole cell extracts were separated by a SDS-PAGE and electroblotted on nitrocellulose membranes. Specific protein detection was carried out with a primary anti-perilipin 1 or anti-perilipin 2 antibody and a horseradish peroxidase labelled anti-rabbit IGG secondary antibody. After visualization of the protein bands by chemiluminescence the perilipin 1 and 2 bands were quantified and normalized to β-actin bands. Shown are means of ± SEM of n = 3 independent immunoblots and one representative blot for each cell line. ^* p < 0.01 compared to control cells without NEFAs (ANOVA/Tukey Multiple Comparison Test). ^# p < 0.01 compared with control cells (ANOVA/Tukey Multiple Comparison Test)