Figure 4. Role of CD36 in PrP_106–126-induced production of nitric oxide and iNOS expression in BV2 and primary microglia.

Cells were first pre-incubated or not with anti-CD36mAb (1 µg/ml) or irrelevant rabbit IgG (Ab) and then treated for 12 hours with PBS, 50 µM PrP_106–126 (PrP), or scrambled PrP_106–126 (Scr). A. The level of NO was measured in supernatant from primary microglia culture with nitrite assay as described in Materials and Methods. The amount of nitric oxide is expressed as U/l cell supernatant. B. ELISA was performed on supernatant from culture of BV2 microglia as described in Materials and methods. All data in A and B are means ± s.d. of triplicate samples and are representative of an experimental n of 3 or 4, * P<0.05. C. Cytoplasmic extracts from primary microglia were prepared and immunoblotted with anti-iNOS antibody as described in Materials and Methods. The blot was stripped and reprobed with anti-β-actin antibody to estimate the total amount of protein loaded in gel. Representative blots of iNOS and actin are shown. Bars represent the relative levels of iNOS, compared with β-actin, and were expressed as arbitrary units. Data are the means±S.D of three independent experiments. * P<0.05, significantly different from control cells. 1-PBS, 2–50 µM PrP_106–126, 3- PrP_106–126+irrelevant rabbit IgG (Ab), 4- Prpscr (50 µM), 5- Anti-CD36mAb (1 µg/ml), 6- PrP_106–126+Anti-CD36mAb.