Fig. 3: Nedd9 depletion enhances the pro-inflammatory response of bacteria-infected macrophages and improves their bacterial clearance ex vivo.

Infection of Nedd9^wt/wt and Nedd9^-/- mBMDMs with ST. A IL-6, unpaired t-test: p = 0.0289 and (B) TNF-α, Mann-Whitney test: p = 0.0286 in supernatants of mBMDMs 6 h post ST infection, n = 4, biological replicates. C Western blot analysis of NFК-B Ser536 phosphorylation 1 and 4 hours post infection with ST and (D) densitometric analysis of pNF-КB Ser536 normalized to GAPDH, n = 4, biological replicates, 2-way ANOVA: p (Nedd9^wt/wt UI vs. ST 1 h) = 0.0005, p (Nedd9^wt/wt UI vs. ST 4 h) = 0.0002, p (Nedd9^wt/wt ST 1 h vs. Nedd9^-/- ST 1 h) = 0.0024, p (Nedd9^wt/wt ST 4 h vs. Nedd9^-/- ST 4 h) = 0.0103. E Western blot analysis of phosphorylated p38 MAPK Tyr180/Tyr182 1 h and 4 h post infection with ST and (F) densitometric analysis of phosphorylated p38 MAPK Tyr180/Tyr182 normalized to GAPDH, n = 2, biological replicates, 2-way ANOVA: p (Nedd9^wt/wt UI vs. ST 1 h) = 0.0090, p (Nedd9^wt/wt ST 1 h vs. Nedd9^-/- ST 1 h) = 0.0043. G In vitro bacterial burden (CFU) in Nedd9^wt/wt vs. Nedd9^-/- mBMDMs plotted as fold reduction of 24 h to uptake (0 h), n = 3, experiments with at least 3 replicates, Mann-Whitney test: p = 0.0286. H CFU of Nedd9^-/- mBMDMs treated with 100 nM Concanamycin A (Conc. A) and controls plotted as fold reduction of 24 h to uptake (0 h), n = 2, experiments with 3 replicates each, unpaired t-test: p = 0.0235. p> 0.05 equals not significant (ns), *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001.