Figure 4: ABCC4 Affects Dense Granule Release and Platelet cAMP Homeostasis in HIV
2019-06-11T07:07:03Z (GMT) by
(A) Mepacrine staining was assessed via flow cytometry in platelets from controls (C) and HIV before and after activation with 0.05 or 0.25 U/ml thrombin. A representative histogram of 1 healthy control is shown. Quantitative analysis of 8 controls and 9 HIV is shown. *p < 0.05 and ***p < 0.0001. (B) Mepacrine staining via flow cytometry of platelets from controls (n = 9) and HIV (n = 8) at basal state. **p< 0.01. (C) Platelets from controls (n = 5) and HIV subjects (n = 4) were pre-treated with Ceefourin 2 (Ceefourin, 10 μmol/l), activated with 0, 0.05, or 0.25 U/ml thrombin and assessed for mepacrine staining. **p < 0.01 and ***p < 0.0001. (D) cAMP was quantified in the supernatants of activated (0.05 U/ml thrombin, 5 min) platelets from controls (n = 9) and subjects with HIV (n = 13). Levels are reported as pg/ml. **p < 0.01. (E) Platelets from HIV (n = 3) and controls (n = 3) were harvested for Western blot analysis. Blots were probed with anti P-VASP Ser 157 and anti-VASP, and loading was controlled with anti-GAPDH. Quantitative analysis was obtained by densitometry using ImageJ 1.51n software (NIH, Bethesda, Maryland). *p < 0.05. cAMP = cyclic adenosine monophosphate; GAPDH = glyceraldehyde 3-phoshate dehydrogenase; MFI = mean fluorescence intensity; VASP = vasodilator-stimulated phosphoprotein; other abbreviations as in Figure 1.