(A–F) Photomicrographs (A, C, E) and quantitation (B, D, F) of the colocalization of mCherry-Mtb and GFP-LC3 (A, B), polyubiquitin (C, D) or LAMP1 (E, F) 15 hr after infection of BMDMs from wild-type or Smurf1−/− mice. Insets show representative mycobacteria that would be considered colocalized with respective markers in wild-type BMDMs or not colocalized in Smurf1−/− BMDMs. Scale bars, 3 μm. See also Figure S1.
(G) Mycobacterial growth in wild-type or Smurf1−/− BMDMs infected with Mtb. Infected cells were lysed at the indicated time-points and mycobacterial growth was determined by counting colony-forming-units (CFUs). Bars are mean ± SEM of quadruplicate samples; each sample was normalized to day 0. Similar results were observed in three independent experiments. ***P<0.01; ****P<0.0001 for indicated comparison; two-way ANOVA.
For B, D and F, bars are mean ± SEM of quadruplicate samples (100 bacteria evaluated per sample per genotype) from a representative experiment. Similar results were observed in at least three independent experiments. *P<0.05, **P<0.01, for indicated comparison; t-test.