CD36 is associated with the FcRγ in human platelets. (A) Washed human platelets (7 × 108/mL), either untreated or treated with oxLDL (50 μg/mL) or nLDL (50 μg/mL) for 15 seconds, were immunoprecipitated for CD36 and immunoblotted for Fyn, Lyn, FcRγ, JNK, and CD36. (Ai) Representative blots (i) and densitometric analysis of FcRγ from 7 independent experiments (ii). (B) Washed human platelets (7 × 108/mL), either untreated or treated with oxLDL (50 μg/mL) or nLDL (50 μg/mL) for 15 seconds, were immunoprecipitated for FcRγ and immunoblotted for CD36. (i) Representative blots and (ii) densitometric analysis of CD36 from 4 independent experiments. (C) Washed human platelets (3 × 106/mL) were adhered to glass coverslips coated with oxLDL (100 μg/mL) and incubated with combinations of CD36 and FcRγ antibodies; CD36 and Lyn antibodies; CD36 and PKA RII antibodies; CD36 antibody alone; FcRγ antibody alone; or antibody diluent alone. Platelets were then stained with Alexa Fluor 488–conjugated wheat germ agglutinin to identify membranes (green) and PLA probes to detect protein-protein interactions (red). Samples were viewed using confocal microscopy under ×63 original magnification. Scale bar, 20 μm. (i) Representative overlaid images and (ii) quantification of PLA/WGA from 4 independent experiments. ∗P < .05, ∗∗∗P < .001. IgG, immunoglobulin G; NS, nonsignificant; WCL, whole-cell lysate; WGA, wheat germ agglutinin.
