Fig. 6: The binding of NEMO with damaging KRT32 mutations. | Nature Communications

Fig. 6: The binding of NEMO with damaging KRT32 mutations.

From: Loss-of-function mutations in Keratin 32 gene disrupt skin immune homeostasis in pityriasis rubra pilaris

Fig. 6

A The molecular docking software HDOCK was used to simulate the interaction between KRT32 and NEMO. B Detailed interactions of KRT32 with NEMO. The confirmed active sites of KRT32 are shown as red sticks, while the other residues critical for binding are shown as blue sticks (KRT32) and golden sticks (NEMO). The hydrogen bond is shown as a green dashed line. C GST-NEMO fusion protein was incubated with E. coli extracts containing WT or six mutant His-KRT32 proteins. GST complexes were isolated with glutathione-agarose beads, and the interacting proteins present in the pull-down eluate were detected by western blotting. D PLA analysis of the interaction of Flag-tagged KRT32 wildtype and mutations with NEMO in Ker-CT cells. Scale bars represent 10 μm. E Quantification of the number of PLA foci per cell in (D). Each dot on the graph corresponds to a specific analyzed cell. Red bars represent the mean ± SEM from the indicated number of cells (N). The number of cells analyzed per group varies as follows: N = 311, 299, 335, 292, 312, 303, 303, and 311, with each group consisting of three biological replicates. P value was calculated using a two-sided unpaired Student’s t test. Source data are provided as Source Data file.

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