4.3 (d) Study of pro-inflammatory cytokine profile in response to Rv2615c
THP-1 macrophages were left unstimulated or stimulated with 10ug/ml of Rv2615c or control- LPS [40ng/ml] for 16h, 24h and 48h were analysed for secretion of TNFα (Thermo Scientific) and IL-1β (Biolegend) in cell culture supernatants. ELISA was done according to manufacturer’s instructions.
ELISA was also performed for estimation of TNFα and IL-1β in supernatants blocked with Anti-TLR4 monoclonal antibody prior to stimulation with Rv2615c/controls till 24 h.