Rv2004c Regulates NF-κB in THP-1 Cells

7 × 10⁶ differentiated THP-1 cells were treated with 1 µg of Rv2004c for 24 h. Untreated cells and cells treated with LPS (100 ng/ml) served as negative and positive controls, respectively. Whole-cell protein from THP-1 cells was prepared according to the protocol described earlier (26 (link)). 100 µg of total protein was subjected to SDS PAGE and transferred onto PVDF membrane. The blots were probed with mouse antihuman p65 NF-κB antibody (Abcam, USA) followed by goat anti-mouse secondary antibody (Sigma Aldrich, USA). Blots were developed in versa doc (Bio-rad) using Super signal west Pico Kit (Thermo Fischer Scientific, USA).