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. 2020 Nov 12;10:19644. doi: 10.1038/s41598-020-76883-9

Figure 2.

Figure 2

The first PDZ motif of ZO-1 protein attenuates LPS-induced F-actin formation. (a) BEAS-2b were treated with LPS (10 µg/ml) in a time-dependent manner. β-actin was used as a loading control. (b) NCI-H292 cells were transfected with either a construct driving the expression of wild-type ZO-1 or ZO-1-specific siRNA. Cells were then incubated with LPS for 10 h. The cells were stained with ActinGreen 488 ReadyProbe reagent. (c) Cells were transfected and were then incubated with LPS for four hours before the generation of total cell lysates, and pro-inflammatory cytokine transcripts were assessed by qRT-PCR. *p < 0.05 compared to the control, **p < 0.05 compared to LPS only, and ***p < 0.05 compared to ZO-1-transfected cells. (d) Constructs were designed according to the amino acids deleted (e.q. M1: 2–156). Black lines represent the magnitude of the sequences encoded by each construct. The expression level of each construct was checked by Western blotting (anti-Flag; right panel). (e) The cells were transfected with either a ZO-1 overexpression construct or deletion constructs and incubated with LPS for 10 h. F-actin was stained with a specific reagent.