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.