Figure 5.
Hemicentin-1 deficiency effect on KIF organization. (A) HaCaT cells grown to 60% confluence on glass coverslips in 12 wells plates were downregulated for HMCN1 with a specific siRNA (siHMCN1) or with a control siRNA (siControl) and co-transfected with an empty vector (EV) or expression vectors encoding either, WT cDNA, or mutant (Mut) KRT14 cDNA carrying the c.373C>T, p.Arg125Cys variant for 48 h at 37°C. The cells were stained for KRT14 expression (red staining) and DAPI (blue staining) (the experiment was repeated two times). Cells were visualized by confocal microscopy. Note protein aggregates in cells transfected with the mutant KRT14 as well as lower expression levels of K14 in siHMCN1-transfected cells (scale bar = 10 μm). (B) K14 KIF fluorescence intensity per cell was measured using ImageJ, analyzing four different zones for each condition. Statistical analysis was performed by t test (***P < 0.005; ****P < 0.001).

Hemicentin-1 deficiency effect on KIF organization. (A) HaCaT cells grown to 60% confluence on glass coverslips in 12 wells plates were downregulated for HMCN1 with a specific siRNA (siHMCN1) or with a control siRNA (siControl) and co-transfected with an empty vector (EV) or expression vectors encoding either, WT cDNA, or mutant (Mut) KRT14 cDNA carrying the c.373C>T, p.Arg125Cys variant for 48 h at 37°C. The cells were stained for KRT14 expression (red staining) and DAPI (blue staining) (the experiment was repeated two times). Cells were visualized by confocal microscopy. Note protein aggregates in cells transfected with the mutant KRT14 as well as lower expression levels of K14 in siHMCN1-transfected cells (scale bar = 10 μm). (B) K14 KIF fluorescence intensity per cell was measured using ImageJ, analyzing four different zones for each condition. Statistical analysis was performed by t test (***P < 0.005; ****P < 0.001).

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