Figure 6

Expression of glycolytic enzymes and HIV-1 Gag in primary CD4+ T cells is not affected by culturing of cells in media containing galactose or glucose. A. Western blot showing the expression of heat shock protein 90 (HSP90), hexokinase 2 (HK2), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), pyruvate kinase M2 (PKM2), pro-caspase 3 (p32), activated caspase 3 (p19) and HIV-1 Gag (p24^Gag and p55^Gag) in HIV-1 infected (NL4.3) and uninfected (Δ Env) primary CD4+ T cells for one representative donor (donor 2). Lanes corresponding to cultures containing galactose are indicated with gal and lanes corresponding to cultures containing glucose are indicated with glc. B. The relative amount of p24^Gag and p55^Gag HIV-1 in primary CD4+ T cells from cultures containing galactose or glucose as quantified from western blots. p24^Gag and p55^Gag were divided by the amount of HSP90 in each sample and normalised to the amount present in cultures containing glucose. The data are the average of a total of five western blots from independent infections of primary CD4+ T cells from three different donors with NL4.3. Errors bars represent the standard deviation. C. The amount of HIV-1 unspliced mRNA relative to β-actin mRNA in primary CD4+ T cells infected with NL4.3 or NL4.3 Δ Env as determined by qPCR. Data are normalised to the amount of HIV-1 Gag mRNA in cultures containing glucose and are the average of five independent infections of primary CD4+ T cells from three different donors. Error bars indicate the standard deviation. D. The ratio of activated caspase 3 p19 to pro-caspase 3 p32 in HIV-1 infected (NL4.3) and uninfected (Δ Env) primary CD4+ T cells from cultures containing galactose or glucose as quantified from western blots.