Fig. 7: Shutdown of Ifih1 rescues the SCs and melanocytes defects in Adar1 mutants.
From: ADAR1 mediated regulation of neural crest derived melanocytes and Schwann cell development
a TUJ1 (green)/MBP (red) immunostaining performed on primary cultures of mixed neurons and SCs from dorsal root ganglia (DRG) of control and HtPA-Cre; Adar1fl/fl mutant embryos at E13.5. Cultures were untransduced (no infection, Ctl) or transduced with lentivirus containing Sh-control (Sh-ctl) or Sh-Ifih1 just before myelination induction. Note the absence of MBP in Adar1 mutants and the restored phenotype upon Sh-Ifih1 transduction. Counterstaining with DAPI (blue) is shown to identify cell density. Scale bar: 12 μm. b Relative mRNA expression of Ifih1 (Mda5, to show Ifih1-shRNAs efficacy validation), and two ISGs (Cxcl10 and Isg15) analyzed by RT-qPCR on RNAs extracted from untransduced (−) and transduced primary cultures of controls (gray) and mutants (blue). All data represent mean ± SEM. Statistical differences between the groups (n = 4 controls and n = 4 mutants) were determined using t test (asterisks represent p values: *p < 0.05, **p < 0.01, ***p < 0.001). c Pigmented melanocytes (melanin-producing cells) observed after culture of NC cells issued from trunk region of neural tube of control and Adar1 mutant embryos at E9.5. No discernable differences could be observed between controls and mutants during the first 7 days of culture, but note the reduction of melanin-producing cells in Adar1 mutants (n = 7) compared to controls (n = 6) after 21 days of culture. Three days after neural tube explant deposition on fibronectin, cultures were untransduced (Ctl) or transduced with lentivirus containing Sh-Ifih1. Note the restored phenotype of mutants upon Sh-Ifih1 transduction (n = 5). Sh-Ifih1 transduction had no discernable effects on control cultures (n = 3). Scale bar: 100 μm. Source data are provided as a source data file.
