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Fig. 4 | Epigenetics & Chromatin

Fig. 4

From: Folate deficiency reduced aberrant level of DOT1L-mediated histone H3K79 methylation causes disruptive SHH gene expression involved in neural tube defects

Fig. 4

MTX-induced downregulation of Shh, Sufu genes expression depends on Dot1l. A siRNA-Dot1l and siRNA-Con were transfected for 24 h in (mouse teratocarcinoma cells) F9 cells, and then cells were treated with MTX for 12 h. Protein was extracted for Western blotting. B, C F9 cells were transfected with siRNA-Dot1l for 24 h, and then treated with MTX (1 μM, 12 h) for immunofluorescence and RT-qPCR detection. D Chromatin immunoprecipitations were performed using digested chromatin from MTX (1 μM, 12 h) treated mESCs and 77,087 Dot1L(D1W4Z)Rabbit mAb. The enrichment of Dot1L at Shh, Sufu genes promoters by RT-qPCR. E F9 cells siRNA-Dot1l and con were transfected for 24 h and treated with MTX for 12 h. Chromatin immunoprecipitations were performed using digested chromatin from F9 cells and H3K79me2, and the enrichment of H3K79me2 at Shh, Sufu genes promoters or intron was detected by RT-qPCR

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Epigenetics & Chromatin

ISSN: 1756-8935