Fig. 6

CpsA is required for the recruitment of galectin-3 to the Mm containing phagosomes followed by release of host and bacterial-derived DNA to the cytosol (A) The levels of mitochondrial (D loop1), nuclear (Tert), or Mm (furA) DNA in the cytosolic compartment of RAW264.7 cells infected with WT or ?cpsA Mm were analyzed by qPCR as described in materials and methods. Data represent means ± SD. ????p < 0.0001 by unpaired t test. (B) The mitochondrial protein TIMM44 and nuclear protein Histone H3 were analyzed in whole-cell lysates (WCL) and cytosolic fractions (CYT) by WB. GAPDH was used as a loading control. (C) Immunofluorescence of RAW264.7 cells infected with Wasabi labeled WT, ?cpsA, the complemented strains, and ?RD1 Mm at MOI of 10 for 4 hpi with uninfected group as control and imaged after staining with anti-galectin-3 (red). Scale bar, 10 ?m. White arrows indicate the colocalization of galectin-3 with bacteria. (D) Quantification of Mm and galectin-3 colocalization in (C). A minimum of 200 cells were quantified per group. The images were representative of three independent experiments. Data represent means ± SD of three independent experiments. ????p < 0.0001 by one-way ANOVA.