Stutter cysteine is sufficient to relocalize YAP1 in differentiating mouse skin keratinocytes. (A) Representative micrographs of Krt14^WT/null keratinocytes in primary culture transfected with EGFP-K14, EGFP-K15, or EGFP-K15 A351C, with and without 1.2 mM CaCl₂ to induce differentiation. YAP1 is visualized through immunofluorescence. “Merge” panels display autofluorescent EGFP (green) and YAP1 (magenta). Arrowheads indicate EGFP-positive keratinocytes. Scale bar, 10 µm. (A′) Magnified insets of nuclei with YAP1 indirect immunofluorescence from Fig. 5 A. Arrowheads indicate EGFP-positive keratinocytes. Scale bar, 10 µm. (B) Scatter plots representing three independent replicates, pooled, displaying mean ± SD. Dots represent a sc. Comparisons were made using unpaired t tests with Welch’s correction. ****P < 0.0001, **P = 0.0058 and 0.0014. (C) Multiplexed immunostaining for K15 (green) and YAP1 (red). Nuclei are counterstained with DAPI (blue). A dashed line specifies the basal lamina. epi, epidermis; derm, dermis. Scale bar, 50 µm. Arrowheads depict region with strong K15 and nuclear YAP1 staining. (D) XY scatter plot is comparing nuclear YAP1 intensity (arbitrary units) vs. K15 intensity (arbitrary units). Each dot represents a single basal keratinocyte. Equation for the simple linear regression of the comparison is displayed, showing a positive correlation. Regression is significantly nonzero (P < 0.0001).