Krt18

Cells were fixed in 4% paraformaldehyde for 20 min, washed with PBS, and permeabilized in 0.5% Triton X-100/PBS for 10 minutes. After blocking with 3% BSA/PBS for 1h, cells were incubated with primary antibodies overnight in 4℃. Cells were then washed in PBS, incubated with the secondary antibodies for 1 hour and washed with PBS three times. Hoechst 33258 (Molecular Probes H1398) was used for nuclei staining (10min). The following primary antibodies were used: anti-TP63 antibody (BA1887, Boster); Cytokeratin 14 (sc-58724, Santa Cruz); Cytokeratin 1 (sc-65999, Santa Cruz); Cytokeratin 10 (sc-23877, Santa Cruz); Involucrin (sc-21748, Santa Cruz); Filaggrin (sc-30229, Santa Cruz); KRT18 (sc-6259, Santa Cruz).

Nuclear and cytoplasmic proteins were extracted according to the manufacturer's instructions (Sigma, #NXTRACT). Western blotting was performed as described in detail in a previous study [26 (link)]. Primary antibodies used in current study included anti-mouse Yap1 (Abcam, #ab56701), TAZ (Cell signaling, #4883), Histone H4 (Santa Cruz, #sc-25260), Hsp90 (Santa Cruz, #sc-8262), p-Smad2 (Santa Cruz, #sc-101801), Smad3 (Cell signaling, #9513), p-Smad3 (Santa Cruz, #sc-11769), Krt14 (Santa Cruz, #sc-43310), Krt18 (Santa Cruz, #sc-45406) and p63 (Abcam, #ab124762).