Fig. 6

Loss-of-Nrf3 in keratinocytes protects from apoptosis by enhancing cell-matrix and cell–cell adhesion. a, b Immortalized keratinocytes from wt and Nrf3-ko mice were treated with tBHQ, sorbitol, accutase, EDTA, or EGTA or irradiated with 10 mJ/cm² UVB. At different time points post treatment they were analyzed by flow cytometry for cleaved caspase-3. c Immortalized mouse keratinocytes were analyzed for adhesion on uncoated plastic or glass dishes or on dishes coated with collagen type I (Col I), type IV (Col IV), or fibronectin (FN) in triplicate wells. Adhesion of wt cells on uncoated dishes was set to 1. d Immortalized keratinocytes were analyzed for detachment from uncoated plastic dishes. Values obtained for non-treated wt cells were set to 100. e Human primary keratinocytes were transfected with NRF3 or CASP5 siRNAs. Cell adhesion was analyzed 24 h after transfection and 24 h after seeding. Adhesion of cells transfected with CASP5 siRNA was set to 1. f Immortalized keratinocytes were analyzed for adhesion on dishes coated with matrix deposited by either wt or Nrf3-ko cells. Adhesion of wt cells on dishes coated with matrix of wt cells was set to 1. g Representative pictures of sheets from wt and Nrf3-ko cells after dispase treatment and exposure to mechanical stress (left) and quantification of fragments (right). Scatter plots with mean and SD are shown. Data points represent results from individual immortalized cell lines derived from different mice or from different siRNA transfection experiments. All results shown are representatives of at least three independent experiments