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. 2018 Dec 3;217(12):4199–4214. doi: 10.1083/jcb.201802155

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© 2018 Singh et al.

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Figure 7. — Role of Syn8 in the survival of Salmonella in host cells. (A) To determine the role of Syn8 in the survival of Salmonella in HeLa cells, Syn8 was knocked down with Syn8 siRNA, and cells were infected with Salmonella:WT as described in Materials and methods. Cells were lysed at the indicated time points after infection, and the number of bacteria present in the cells was calculated by measuring colony-forming units. Results are expressed as percent survival, arbitrarily chosen as 100% survival at 0 h of infection with Salmonella in control cells. All results are mean ± SEM of three independent experiments, and levels of significance are indicated by P values. Inset shows the specific knockdown of Syn8 by siRNA in HeLa cells by qPCR. (B) A similar assay was performed in CRISPR-mediated KO of Syn8 in HeLa cells. Inset shows the specific KO of Syn8 by CRISPR in HeLa cells by immunofluorescence and Western blot analysis using specific antibodies. (C) Lysosomes of HeLa cells were labeled with Dextran–Texas Red and were infected with GFP-Salmonella:WT or GFP-Salmonella:sipAKO. Cells were analyzed by confocal microscopy at indicated time points to determine the localization of respective Salmonella in the lysosomes. Bottom: Quantitation for the same. P values are indicated.