FIG. 1.

Targeted siRNA knockdown of kinases in PBMC resulting impact on FTC-MP, FTC-DP and FTC-TP intracellular formation. PBMC were electroporated with 500 nM nontargeting siRNA or siRNA targeting DCK, TK1, CMPK1, or PGK1 and incubated for 48 h. Representative immunoblots demonstrate decreased kinase expression with each targeted siRNA treatment relative to the nontargeting siRNA control. siRNA treated PBMC were incubated with 10 μM FTC for 12 h (n = 5). Intracellular anabolites were extracted and FTC-MP, FTC-DP, and FTC-TP were measured using uHPLC-UV as depicted in the corresponding bar graphs showing mean ± standard deviation for each treatment. Statistical analyses were performed using a two-tailed unpaired t test to compare relative levels of anabolite production between nontargeted and targeted siRNA conditions; **p ≤ 0.01; ***p ≤ 0.001. FTC, fumarate and emtricitabine; PBMC, peripheral blood mononuclear cell.