Figure 7
From: Secretory phospholipase A2 modified HDL rapidly and potently suppresses platelet activation
sPLA2-HDL inhibits Akt and ERK1/2 phosphorylation in platelets. Platelets were pretreated with vehicle, nHDL (50 µg/mL), sPLA2-HDL-low (50 µg/mL) or sPLA2-HDL-high (50 µg/mL). Subsequently cells were stimulated with (a,c) ADP (10 µM) or (b,d) collagen (5 µg/mL). (a,b) Akt (Ser473) and (c,d) ERK1/2 phosphorylation was assessed by Western blot. β-actin was used as the loading control. Blots have been cropped from full-length blots shown in Supplementary Figure 4. ChemiDoc Touch Imaging System and ECL Blotting Substrate (both Bio-Rad, Vienna, Austria) were used to visualize protein bands. Immunoblot images were quantified using Image Lab 5.2 software (Bio-Rad). Statistical analysis of phospho-Akt (p-Akt)/total Akt (t-Akt) and phospho-ERK1/2 (p-ERK)/total ERK1/2 (t-ERK) ratios were derived from 3 independent experiments which were run under the same conditions. Vehicle treated (unstimulated) control was set as baseline and values are expressed as % over baseline. Results are shown as mean ± SEM. Statistical significance was assessed by one-way ANOVA followed by Dunnett’s post hoc test. *p < 0.05, **p < 0.01 versus vehicle-pretreated and (a,c) ADP or (b,d) collagen-stimulated platelets.
