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. 2021 Jun 4;28(11):3161–3175. doi: 10.1038/s41418-021-00808-w

Fig. 6. RNF20 and RNF40 deficiency results in reduced VDR levels.

Fig. 6

A Rnf20 and Rnf40 deletion reduced Vdr mRNA and (B) protein levels in IECs as demonstrated by qRT-PCR and Western blot. C IECs were isolated from wild type mice and cultured under vitamin D-deficient conditions. RNF20 and RNF40 were transiently depleted using siRNA and 24 h after knockdown cells were treated with calcitriol (1 µM) for 48 h (n = 3). D Calcitriol treatment induced the mRNA expression of VDR target genes in control cells but not in RNF20- and RNF40-depleted IECs as detected using qRT-PCR. E The reduction of Vdr following Rnf20 and Rnf40 genetic deletion was confirmed in whole colon lysates using qRT-PCR (n = 6). F IHC for VDR and subsequent (G) determination of staining intensity using FIJI verified reduced VDR protein amounts in Rnf20fl/fl and Rnf40fl/fl colons. Scale bar: 100 µm. One-way ANOVA, mean ± SEM.