NRF2 and its target genes are expressed during Mtb infection.

(A) BALB/c mouse peritoneal macrophages were infected with H37Rv for 48 h and the expression of NRF2 target genes was assessed by qRT-PCR. (B) BALB/c mouse peritoneal macrophages were infected with H37Rv for the indicated time points and whole cell lysates were assessed for the expression of NRF2 and its target genes. (C) Immunoblotting to validate NRF2 knockdown in murine macrophages transfected with Nfe2l2 siRNA. (D-F) BALB/c mouse peritoneal macrophages were transfected with NT or (D) Nfe2l2 siRNA or (E) Ehmt2 and Sirt6 siRNA or Chol accum genes siRNA (combination of Lrp2, Aacs, Hmgcs1, Mvd and Dhcr24 siRNAs) or (F) siRNAs against the selected cholesterol accumulation genes (combination of Lrp2, Aacs, Hmgcs1, Mvd and Dhcr24 siRNAs) or NT and assessed for the indicated transcript by qRT-PCR. (G) MTT assay was performed to assess cell viability of BALB/c macrophages transfected with NT or siRNA against selected cholesterol accumulation genes. (H-I) BALB/c mouse peritoneal macrophages were infected with H37Rv for 48 h and stained with CellROX to assess for oxidative stress was performed. Representative images and (H) its quantification (I). (J) BALB/c mouse peritoneal macrophages were transfected with siRNAs against the selected cholesterol accumulation genes (combination of Lrp2, Aacs, Hmgcs1, Mvd and Dhcr24 siRNAs) or NT and followed by H37Rv infection for 48 h and cholesterol accumulation was confirmed by Filipin staining;. The MOI of infection is 1:10 (macrophage:mycobacteria) for all the in vitro experiments. All data represents the mean ± SEM from 3 independent experiments; *, P (TIF)