Figure 2

Phagocytosis and survival of BCG in PKC-α deficient THP-1 cells. THP-1 cells were incubated in the presence of 30 nM PMA for 24 h. Then cells were transfected with 20 nM SiRNA and level of PKC-α were determined by immunoblotting. (A) 24 h after transfection, level of PKC-α and PKC-δ in cells transfected with SiRNA targeting PKC-α or scrambled SiRNA, (B) 24 h after transfection, (ΔA) cells transfected with SiRNA targeting PKC-α and (S) cells transfected with scrambled SiRNA and control cells (C) were infected with BCG (MOI = 1:10) for 2 h, washed and remaining extracellular bacilli were killed by amikacin treatment for 1 h and lysed in 0.05% SDS and plated. Colony forming units (cfu) were determined after 4 week of incubation. Tukey (T) test was performed for statistical analysis of data (C) Survival of BCG in THP-1 cells transfected with either SiRNA targeting PKC-α (ΔA) or scrambled SiRNA (S) after 24 and 48 h, since phagocytosis of BCG in control and PKC-α deficient cells was different, CFU at 0 h was considered 1 and survival of BCG is presented as fold increase in the number of cfu as compared to the initial phagocytosis. Data are means ± standard deviations from three independent experiments each performed in 4 replicates. (** = p < 0.005).