Figure 5.

Rv2145c induces activation of STAT3 in Mtb-infected macrophages. (A, B) BMDMs were infected with (A) Mtb at an MOI of 1 for 4 h and incubated with or without 10 μg/ml Rv2145c or (B) Ms_vector or Ms_Rv2145c at an MOI of 10 for 4 h. The cells were lysed at the indicated times, and the proteins in the total cell lysate were separated by SDS–PAGE, followed by immunoblot analysis using antibodies against phospho-STAT3, STAT3, phospho-STAT1, STAT1, iNOs and β-actin. This image is representative of three experiments showing similar results. (C, D) At 48 h after Mtb infection (C) or at 24 h after M. smegmatis infection (D), BMDMs were fixed with 4% paraformaldehyde and immunolabeled with an anti-phospho-STAT3 antibody, followed by Alexa 488-conjugated goat anti-rabbit IgG. The cells were stained with DAPI to visualize the nuclei (blue). The localization of the target molecules was analyzed by laser-scanning confocal microscopy. Scale bar, 10 μm. MC, medium controls.