Fig. 1
SNAIL1 expression controls fibronectin EDA inclusion. A Relative RNA amount of EDA+ fibronectin isoforms in control and Snai1 KO CAFs. RNA obtained from indicated CAFs was retrotranscribed and amplified by PCR using Fn1 (depicted on the left) or Snai1 primers. Resulting DNA was visualized by electrophoresis on a 2% agarose gel. A representative experiment from the three performed is shown. B Relative RNA amount of EDA+ fibronectin isoforms in control and Snai1 KO MEFs treated with TGFβ1. RNA from the indicated MEFs was untreated or treated for 24 h with 5 ng/mL of TGFβ1 and then analyzed as in A. C Expression of the EDA+ fibronectin protein in MEFs. Control and Snai1 KO MEFs were treated or not with 5 ng/mL of TGFβ1 for 24 h and lysed in SDS buffer. Levels of the indicated proteins were analyzed by Western blotting. D Expression of EDA+ fibronectin in BJ fibroblasts. Human BJ fibroblasts were transfected with siRNA anti-SNAIL1 or a control siRNA and then treated or not with TGFβ1 for 24 h. Cells were lysed in SDS buffer, and protein levels were analyzed by Western blotting. E Quantification of the EDA+ fibronectin ratio by RNA-seq. Deep sequencing of RNA from control or Snai1 KO MEFs treated with 5 ng/mL TGFβ1 for 3 h was performed. The percent spliced-in (PSI) for FN1-EDA in each condition was calculated from the number of inclusion and exclusion sequencing reads