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. 2020 Sep 24;16(9):e1009040. doi: 10.1371/journal.pgen.1009040

Fig 4. Inhibition of RA signaling upregulates Pou4f3 expression in cochlear explants.

Fig 4

(A) Upregulation of Pou4f3 mRNA expression by DEAB and BMS195614 in P3 wildtype cochlear explants. DEAB and BMS195614 (BMS) concentrations were 100 and 5 μM, respectively. DMSO was used as control (-). * P < 0.05 and ** P < 0.01 by unpaired student’s t-test, n = 13–15 explants for DEAB, n = 4–8 explants for BMS195614. (B) Effects of DEAB (red bar) on mRNA expressions of Tmem237 and Sra1. * P < 0.05 by unpaired student’s t-test, n = 4 from each treatment. (C) Effects of DEAB (red bar) on mRNA expressions of known Pou4f3 targets. * P < 0.05 and *** P < 0.001 by unpaired student’s t-test, n = 4 from each treatment. (D-F) Effect of DEAB (100 μM) and RA (0.3 μM) treatment on mRNA expressions of (D) Cyp26a1, (E) Pou4f3 and (F) Lhx3 in P3 wildtype cochlear explants. DMSO was used as control (-). * P < 0.05, ** P < 0.01 and *** P < 0.001 by one-way ANOVA, n = 4 explants for each treatment. (G) Upregulation of Pou4f3 mRNA expression by DEAB in P3 Pou4f3(Δ/+) cochlear explants. ** P < 0.01 by unpaired student’s t-test, n = 6 explants for each treatment. (H) Western blots and (I) densitometrical measurements show upregulation of wildtype Pou4f3 protein expression by DEAB in Pou4f3(Δ/+) cochlear explants. The black open arrow indicates the wildtype Pou4f3 and red open arrow shows the truncated mutant Pou4f3 protein. * P < 0.05 by unpaired student’s t-test, n = 4 explants from each treatment.