Figure 3.
MMP10 and S100a7a15 are novel transcriptional targets of c-Fos. (A) Schematic representation of the two most up-regulated genes in a microarray expression analysis comparing in vitro cultured c-FostetON [coltetO-Fos (+/KI), Rosa-rtTA (+/KI)] keratinocytes treated ±Dox (1 mg/mL; n = 3) for 6, 12, 24, 48, and 72 h. (B) RT-qPCR expression analyses of c-fos, mmp10, and s100a7a15 mRNA in c-FostetON keratinocytes treated ±Dox (1 μg/mL; n = 3). (C) Immunoblot depicting S100a7a15 and MMP10 protein levels as well as vinculin and GAPDH as loading controls in c-FostetON keratinocytes treated ±Dox for 48 h. (D) RT-qPCR expression analyses of c-fos, mmp10, and s100a7a15 mRNA in control and c-FosEp-tetON back skin at 4 wk of inducible c-fos expression (n = 3). Mean ± SD. (E) MMP10 immunohistochemical analyses of the back skin of control and c-FosEp-tetON mice after 2 and 4 wk of inducible c-fos expression (n = 3). (F) S100a7a15 immunofluorescence analyses in the back skin of control and c-FosEp-tetON mice after 4 wk of inducible c-fos expression (n = 3). (G) ChIP of c-Fos at the mmp10 promoter. (Left, top) Scheme of the TRE element at the mmp10 promoter where c-Fos binds. (Right) Endpoint qPCR fragments are shown together with the representation of the percentage of binding of c-Fos to mmp10 promoter. (Left, bottom) Chromatin was immunoprecipitated using c-Fos antibody from c-FostetON keratinocytes treated ±Dox for 24 h. (H) ChIP of c-Fos at the s100a7a15 promoter. (Left, top) Scheme of the TRE element at the mmp10 promoter where c-Fos binds. (Right) Endpoint qPCR-fragments are shown together with the representation of the percentage of binding of c-Fos to s100a7a15 promoters. (Left, bottom) Chromatin was immunoprecipitated using c-Fos antibody from c-FostetON keratinocytes treated ±Dox for 24 h.