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. 2022 Apr 1;13:823685. doi: 10.3389/fimmu.2022.823685

Figure 6.

Figure 6

Fimbriae purified from the WT-ATCC 33277 strain stimulate PHGFs to produce cytokines and activate the NF-ĸB and MAPK kinase signaling pathways. (A, B) Western blot analysis of phosphorylated and total p65 and p38 in PHGFs (n=7) treated for 1 h with purified fimbriae (10 µg/ml) isolated from WT-ATCC 33277 (FimA WT) or its isogenic PPAD mutant (FimA delPPAD). β-actin was used as a control. (A) Representative blots and (B) results of densitometry analysis are shown. (C) Expression of mRNA encoding PGE2 synthesis-related genes COX-2 and mPGES-1 in PHGFs (n=7) treated for 4 h with purified fimbriae (10 µg/ml) isolated from WT-ATCC 33277 strain (FimA WT) or its isogenic PPAD mutant (FimA delPPAD). (D, E) Secretion of IL-6 (D) and IL-8 (E) by PHGFs (n=5) treated for 24 h with purified fimbriae (10 µg/ml) from WT and delPPAD ATCC 33277 strains. Data represent the mean ± SEM normalized phosphorylated to total protein ratio (B), relative mRNA expression (C) or fold change of cytokine concentration (D, E); ***p < 0.001; **p < 0.01; *p < 0.05; ns, no statistical significance; 1-way ANOVA followed by Tukey’s multiple comparisons test; “n” represents the number of independent experiments performed on PHGFs cell lines derived from different healthy donors.