Decrease of SMAD3 protein, but not SMAD2 in DBA iPSCs.

<p>DBA iPSCs, the corrected DBA cells, and the control GFP cells were cultured, and the RNA, total protein, protein from nuclear fraction were extracted from iPSCs. The ratio of protein level to WT is labelled under the image. A) No change of mRNA level of <i>SMAD2</i> or <i>SMAD3</i> in the DBA iPSCs with <i>RPS19</i> or <i>RPL5</i> mutations. B) Western blot analysis of total protein showing some decrease in SMAD3 protein but the change of SMAD2 was not obvious. C) Western blot analysis of nuclear protein showing decrease in SMAD3 protein in both DBA lines, and no change in SMAD2 protein. D) Western blot analysis showing slight change of p-SMAD2 level in the nuclear fractions from <i>RPS19</i> mutant compared to WT and the corrected lines. The nuclear p-SMAD2 decreased in <i>RPL5</i> mutant and corrected lines. E) Western blot showing the decrease of p-SMAD3 in the nuclear fraction in both DBA iPS lines, and the nuclear p-SMAD3 protein partially restored in the corrected DBA iPSCs in the <i>RPS19</i> corrected line, but not in <i>RPL5</i> corrected line.