Figure 2

Overexpression of bCoronin-1A facilitated M.tb evasion of lysosomal clearance. A Immunofluorescence detection of M.tb localization after 4 h of infection, with FITC-labeled H37Ra/BCG (green), rabbit anti-LAMP1 monoclonal antibody for lysosomes (red), and DAPI-stained nuclei (blue) (MOI = 20). B Statistical comparison of percent co-localization of M.tb with lysosomes. C Flow cytometry determination of the percentage of extracted lysosomes containing FITC-labeled M.tb (MOI = 10). D The degree of lysosomal fusion in extracted phagosomes was determined by western blot with mouse anti-EEA1 monoclonal antibody for phagosomes and rabbit anti-LAMP1 monoclonal antibody for lysosomes (MOI = 10). E Lysosomal acidification was detected using the LysoSensor™ yellow/blue probe, which produces blue fluorescence under neutral conditions and emits yellow fluorescence when the environment is acidic (MOI = 10). F The intensity of blue and yellow fluorescence was measured with a microplate reader. G The relative mRNA expression of lysosome-related genes was determined by qRT-PCR (MOI = 10). Data were analyzed by t-test and presented as the mean ± SEM of three independent experiments. * P < 0.05, ** P < 0.01, *** P < 0.001, ^nsP > 0.05.