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. 2018 May 18;145(10):dev153619. doi: 10.1242/dev.153619

Fig. 10.

Fig. 10.

Arrested RV differentiation in HDAC1/2 mutant kidneys is Wnt4 independent. (A-C) Section ISH shows maintained expression of Wnt4 in the mutant RVs at E16.5 (A,B) and enhanced total kidney Wnt4 by RT-qPCR (C). *P<0.05; n=4. (D,E) Section IF at P0 using antibodies against Lef1. In WT kidneys, Lef1 is expressed in RVs (arrowheads) and in a thin layer of stromal cells surrounding the UB branches. In HDAC1/2 mutant kidneys, Lef1 is ectopically expressed in the stroma (small arrows) and is upregulated in the peri-UB stroma. (F) FACS-isolated NPCs from WT (n+3) and HDAC1/2 mutant (n=2) were cultured in differentiation medium (with 3.5 µM CHIR) for 48 h. ddPCR showed that the Lef1 RNA copy number of mutant NPCs is not significantly different from that of WT cells, whereas the Six2 RNA copy number of mutant NPC is significantly lower than that of WT cells. Data are mean±s.e.m.