Fig. 9

Intracellular ATP production and intracellular lactate levels and release in PBLs expressing Tat101. Resting PBLs were transiently transfected with vectors CMV-Tat101 or pcDNA3, as negative control, along with pEGFP expression vector, as control of transfection efficiency. a Flow cytometry quantification of the percentage of living cells expressing EGFP was used to analyze transfection efficiency. Intracellular expression of Tat and nuclear subcellular localization were confirmed by immunofluorescence using a monoclonal antibody against Tat and a secondary antibody conjugated to Alexa 546. DAPI was used for nuclear staining. b ATP production was measured by chemiluminescence using a commercial assay. Data shown are RLUs mean and SEM of concentration from five independent experiments. c Relative intracellular lactate production measured in PBLs expressing Tat versus control PBLs. Data shown are relative mean and SEM of concentration from five independent experiments. d Relative release of lactate to the culture medium measured in the same PBLs expressing Tat used to measure intracellular lactate. Statistical significance was calculated by Mann–Whitney test (*p < 0.05)