Figure 3

Effect of HIV1 p55-gag and REV on MSC osteogenesis. MSCs were cultured in a 96-well plate format and induced to differentiate in the presence and absence of HIV REV and p55-gag. Untreated, undifferentiated cells were included as controls. Cells were harvested at 3, 6, 9 and 15 days post induction of differentiation, fixed and stained for either calcium deposition (alizarin red) or alkaline phosphatase (BCIP/NBT). Plates were first photographed, then the stain extracted, and the degree of staining quantified spectrophotometrically and normalised to cell number. Fig 3a; alkaline phosphatase (ALP) staining in untreated MSCs (con), diferentiating MSCs (cd), and differentiating MSCs exposed to HIV p55-gag and REV proteins. Fig 3b; ALP staining normalised to cell number, data expressed as percentage of untreated control ± SEM. Fig 3c; calcium staining in untreated MSCs (con), diferentiating MSCs (cd), and differentiating MSCs exposed to HIV p55-gag and REV proteins. Fig 3d; calcium staining normalised to cell number, data expressed as percentage of untreated control ± SEM. All data is the mean of several independent experiments. (Significance determined at; *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.005 relative to undifferentiated control, ^p ≤ 0.05, ^^p ≤ 0.01, ^^^p ≤ 0.005 relative to differentiated control).