2.2.2 (a) Rv2615c protein induces cell death in THP-1 macrophages
To determine the cell viability, MTT and CellTiter Blue assay were performed to probe any change in viability of THP-1 cells in response to Rv2615c stimulation. Three concentrations of Rv2615c (5μg/ml, 10μg/ml and 15μ/ml) were included for checking the dose dependent viability. Cell viability was prominently affected in Rv2615c stimulated cells in a dose dependent manner (Fig S2a). Based on these observations, we selected 10μg/ml as stimulation concentration for Rv2615c protein in our experiments. Unless otherwise stated, LPS and staurosporine served as controls in each experiment. Viability of Rv2615c-stimulated and control-stimulated THP-1 macrophages was significantly decreased at all-time points when compared to unstimulated cells (Fig S2b and S2c). In comparison to the unstimulated cells, Rv2615c-stimulation caused approximately 50 to 60% cell death till 48 hours that was comparable to cell death induced by LPS stimulation (Fig S2b and S2c). These observations indicate that Rv2615c protein affects viability and induce cell death in THP-1 macrophages.