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. 2018 May 14;8:146. doi: 10.3389/fcimb.2018.00146

Figure 6.

Figure 6

K8 and K18 depletion perturbs expression and surface localization of transmembrane receptors. (A) Surface proteins of control (Ctr), K8- (K8-si), and K18-depleted (K18-si) HeLa cells were biotinylated, recovered from total cell extracts and pulled down using neutravidin beads. Biotinylated samples, which corresponds to surface exposed proteins, and whole cell lysates (WCL) were immunoblotted to detect cMet, TfR, and integrin β1, together with Actin, K8, and K18. (B) Quantifications of TfR in WCL (left panel) and in biotinylated samples (right panel) from at least three independent experiments. (C) Quantifications of integrin β1 in WCL (left panel) and in biotinylated samples (right panel) from at least three independent experiments (a.u., arbitrary units). (D) Functional impact of K18 in the expression of ITGB1 was assessed by gentamicin survival assay and CFU counting in K18-depleted HeLa cells (K18-si) incubated with invasive E. coli K12 expressing the Y. pseudotuberculosis invasin (K12-inv). Values of intracellular bacteria in Ctr cells were normalized to 100% and the entry levels in K18-si cells are expressed as relative values. Values are the mean ± S.E. of three independent experiments, each done in triplicate. Statistically significant differences are indicated: **p < 0.01, ***p < 0.001, and ****p < 0.0001.