Figure 4. Interaction of SNX10 and caspase‐5 is essential for OMV‐induced Lyn phosphorylation.

1. Lysates from WT and SNX10 KO Caco‐2 cells treated with or without 100 μg/ml OMVs for 24 h were analyzed by immunoblots with the indicated antibodies.
2. Lysates from Caco‐2 cells transfected with SNX10‐Flag were subjected to pull‐down assay with anti‐Flag antibody‐conjugated agarose, followed by immunoblots with the indicated antibodies.
3. Representative images of co‐staining of SNX10‐Flag and caspase‐5 in Caco‐2 cells expressing SNX10‐Flag. Scale bar: 20 μm.
4. WT and SNX10 KO Caco‐2 cells were treated with or without OMVs (100 μg/ml) for 24 h and then subjected to IP with anti‐caspase‐5 antibody.
5. Representative images of costaining of Lyn and caspase‐5 in WT and SNX10 KO Caco‐2 cells treated with or without OMVs (100 μg/ml) for 24 h. Scale bar: 20 μm.
6. The co‐localization of Lyn and caspase‐5 was quantified by ImageJ software. n = 3 independent experiments, n = 18 fields analyzed. Data are means ± SD. One‐way ANOVA followed by Bonferroni post hoc test was used for statistical analyses. ***P < 0.001.
7. Cytoplasmic and nuclear proteins were extracted from Caco‐2 cells transfected with control siRNA, CASP4 siRNA, or CASP5 siRNA and the indicated proteins were detected by immunoblots.
8. Lysates of Flag‐tagged full‐length SNX10 or its different truncated mutants transfected with Caco‐2 cells were subjected to immunoprecipitation.
9. Caco‐2 cells were transfected with Flag‐tagged full‐length SNX10 or its indicated truncated mutant, followed by treatment with or without OMVs (100 μg/ml) for 24 h. Lysates were extracted and subjected to immunoblots with the indicated antibodies.

Source data are available online for this figure.