Figure 4. HIV ssRNA40 mediated activation of microglia can induce cytotoxicity and cell death in human primary neurons.

Cell culture supernatants were collected from ssRNA40, ssRNA41 and vehicle treated HMG cells at 48h. HPN cells were cultured in 10% (v/v) neurons culture medium mixed with supernatants derived from HMG cells for 24h at 37°C. (a) LDH release in HPN culture supernatants at 24h post incubation with HMG derived supernatants. 10% neuronal culture medium was used for background normalization. (b) Cell death analysis in HPN cells incubated with 10% HMG derived culture supernatants using ELISA based determination of nucleosomal DNA in the cytoplasmic fraction. Data are presented as mean ± SD, n=4; *P<0.05. (c) Representative immunofluorescent images of HPN cells treated with 10% culture supernatants from HMG cells and stained with MAP2 (green) and DAPI (blue) showing a decrease in HPN number and neurite length. Scale bar indicates 20µm in 20X view and 10µm in 60X view (n=2).