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. 2023 Feb 19;10(12):2205272. doi: 10.1002/advs.202205272

Figure 5.

Figure 5

Active T6SS in K. variicola influences the expression of ZO‐1. A) Components of T6SS in K. variicola ZSLY01 based on data derived from whole‐genome sequencing. B,C) Contact inhibition of K. variicola ZSLY01 (white colony) toward E. coli LF82 (red colony). D) Schematic diagram illustrating the knockdown of ClpV by CRISPR interference (CRISPRi) system, and the mRNA expression of ClpV is significantly inhibited in K.v‐dcas‐ClpV compared to K.v‐vector. E) The contact inhibition effect toward E. coli LF82 was significantly attenuated in K.v‐dcas‐ClpV compared to K.v‐vector. F) The growth rate curves of K.v‐dcas‐ClpV and K.v‐vector. G,H) ZO‐1 immunofluorescent staining and quantification of K.v‐dcas‐ClpV and K.v‐vector treated IEC‐6. I,J) The pro‐inflammatory effects of K.v‐dcas‐ClpV and K.v‐vector toward IEC‐6 and 3T3L1. Bacteria‐cell co‐culture assay: MOI = 20, treated for 4 h. K.v‐vector: K. variicola ZSLY01 transformed with empty plasmid; K.v‐dcas‐ClpV, K. variicola ZSLY01 transformated with sgRNA containing plasmid. Error bars ± SEM. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001; Mann–Whitney U‐test. Results are representative of three independent experiments.