Figure 4: iRHOM2 regulates the reorganization and dynamicity of K16 filaments.
From: Rhomboid family member 2 regulates cytoskeletal stress-associated Keratin 16
(a) CTRL keratinocytes were transfected with either WT-iRHOM2-GFP or TOC Mutant-iRHOM2-GFP and immunostained for endogenous K16 and analysed by confocal microscopy. Scale bar, 20 μm. (b) CTRL and TOC keratinocytes were subjected to cyclical mechanical stretch at a frequency of 5 Hz and amplitude 10–13% using Flexcell FX-4000 Tension system for 0 and 4 h, respectively. Stretched cells were immunostained for K16 and analysed by confocal microscopy. White arrowheads indicate perinuclear K16 filaments reorganization. Scale bar, 20 μm. (c) Quantification of the perinuclear localization of K16 filaments in CTRL and TOC cells. Data represent the mean of 3 independent experiments, each experiment comprising 30 images per cell line. Error bars denote s.d. **P<0.01 (Student’s t-test). (d) Confocal analysis of CTRL and TOC keratinocytes treated with 1 μM OA for 1 h, then fixed 4 and 24 h following treatment and immunostained for K16. White arrowheads represent aggregated K16 filaments. Scale bar, 20 μm.
