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. 2024 Apr 10;27(5):109623. doi: 10.1016/j.isci.2024.109623

Figure 5.

Figure 5

ULK1 is involved in bacterial invasion regulated by ATG9B

(A) WT and ATG9AB KO HeLa cells were treated with the indicated siRNA and infected with GAS at an MOI of 10. The relative percent internalization was normalized with data from non-targeting control siRNA-treated cells (colony-forming units [CFU] recovered at 2 h post-infection [hpi]/CFUs at 1 hpi).

(B–D) Immunostaining of the indicated siRNA-treated, fixed HeLa cells for p-Paxillin (green) and F-actin (magenta). Cellular DNA was stained with DAPI (cyan). Asterisks indicate actin aggregation. Representative microscopic images (B) and quantification of the number of p-Paxillin particles (C) and percentage of cells with F-actin aggregation.€.

(E) Co-IP between FLAG-tagged ATG9B and mClover-tagged ULK1 or ULK1 kinase inactive mutant (K46I) proteins.

(F) Schematic representation of the cytoplasmic domain (CD) mutants of ATG9B.

(G) Co-IP between the indicated FLAG-tagged ATG9B and mClover-tagged ULK1 KI mutant proteins.Data in c (n = 200 cells per condition) and d (n = 100 cells per condition) represent the mean ± SEM from five independent experiments. Scale bar, 10 μm. One-way ANOVA was performed and p values were calculated using Tukey’s multiple comparison test; ∗p < 0.05, ∗∗p < 0.01.