Fig. 2: The effects of CD82 on cytokine production and bacterial survival in macrophages.

BMDMs were infected with MTB Rv or MTB Ra for the indicated times, followed by a immunoblotting (IB) with αCD82 and αActin, or b culture supernatants were harvested and analyzed for cytokine ELISA for TNFα, IL-6, and IL-12p40. BMDMs were transduced with lentivirus-shRNA-NS or lentivirus-shRNA-CD82 for 2 days (c, d) or BMDMs from CD82^+/+ and CD82^−/− (f, g) were with MTB Rv or MTB Ra for the indicated times, followed by intracellular survival of MTB assessed by CFU assay (c, f), or culture supernatants were harvested at 18 h and analyzed for cytokine ELISA for TNFα, IL-6, and IL-12p40 (d, g). e BMDMs from CD82^+/+ and CD82^−/− were infected with MTB Rv or MTB Ra for 6 h. Mycobacteria-containing phagosome fractions were subsequently purified by sucrose-step-gradient ultra-centrifugation, followed by IB to detect αCD82, αRab5, αRab22, αRab7, αLAMP1, αLAMP2, and αActin. The polyclonal MTB Ab and LpqH Ab detect the lipoglycans (LAM and LM) and lipoproteins (LpqH), respectively. The data are representative of four independent experiments with similar results (a, e). Data shown are the means ± SD of six experiments (b–d, f, g). Significant differences (*P < 0.05; **P < 0.01; ***P < 0.001) compared with shRNA-NS or CD82^+/+ (Student’s t-test with Bonferroni adjustment). CFU colony-forming units