FIGURE 4.

JMJD1C knockdown reduces miR-302 expression, enhances TGFβ signaling, and inhibits BMP signaling. A, RT-PCR analysis for expression of pri-miR-302 in JMJD1C KD and MM cells from both H9 and CT2 hESCs. B, Northern blotting for miR-302 isoforms in the KD and MM cells. The small nuclear RNA U6 was determined as a loading control. C, RT-PCR analysis for expression of some TGFβ ligands and targets in wild-type, JMJD1C KD, and MM H9 cells. D, Western blot analysis for phosphorylated SMAD1/5/8 (p-SMAD1/5/8) and phosphorylated SMAD2/3 (p-SMAD2/3) to determine BMP and TGFβ signaling levels, respectively, in the JMJD1C KD and MM (CT2 and H9) cells. Total SMAD1/5/8 and SMAD2/3 were detected as controls.