No direct relationship between KRT17 and stratifin (SFN).
(A) Western blot analysis of Ca9/17+ (C-1, C-2, C-3, and C-4), Ca9, HSC3/K17- (Kd-1, Kd-2, Kd-3, and Kd-4) and HSC3 for detecting SFN and GAPDH. (B) SFN was detected only in the cytoplasmic fractions of the cell lysates. Ca9, Ca9/K17+ (C-4), HSC3 and HSC3/K17- (Kd-4) were lysed and cytoplasmic and nuclear protein fractions were separately extracted. Western blot analysis for detecting SFN, GAPDH and HISTH3. Nuclear translocation or change in the expression level of SFN was not observed. Representative graphs of two assays that showed similar results. (C) Serum starvation did not alter the cytoplasmic localization of SFN. After serum starvation for 12 h, Ca9 and HSC3 cells were lysed and subjected to SDS-PAGE followed by western blot analysis. (D) Absence of KRT13 or KRT17 coprecipitation in SFN immunoprecipitate. 293T cell were transfected by the indicated combinations of plasmids and immunoprecipitation was performed using anti-Flag antibody followed by western blot analysis using an anti-HA antibody. (E) Cytoplasmic localization of SFN in HSC3 and HSC3-KO. Immunofluorescent images showing cytoplasmic staining of SFN (green) counterstained with DAPI (blue).