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. 2018 May 7;19(6):e45657. doi: 10.15252/embr.201745657

Figure 7. Mbnl1/2 bind and promote the expression of key miR‐294 targets.

Figure 7

  1. RIP‐qPCR analysis of key miR‐294 targets in regulating proliferation, apoptosis, and EMT. ESCs were harvested ˜48 h after overexpression. Data were normalized to β‐actin and then to RNA pulled down in GFP overexpressing ESCs. Shown are mean ± SD, n = 3. P values are as indicated. Unpaired two‐tailed Student's t‐test.
  2. RT‐qPCR analysis of key miR‐294 targets in regulating proliferation, apoptosis, and EMT. ESCs were harvested ˜48 h after overexpression. Data were normalized to β‐actin and then to GFP overexpressing ESCs. Shown are mean ± SD, n = 3. P values are as indicated. Unpaired two‐tailed Student's t‐test.
  3. Luciferase assay for 3′UTRs of Cdkn1a and Lats2. The 3′UTRs of Cdkn1a and Lats2 were cloned in pGL3‐basic vectors. Renilla was used as transfection control. Data were normalized to empty vectors and then to GFP overexpressing ESCs. Shown are mean ± SD, n = 4. P values are as indicated. Unpaired two‐tailed Student's t‐test.
  4. Luciferase assay for 3′UTRs of Rhoc, Tgfbr2, and Vim. The 3′UTRs of Rhoc, Tgfbr2, and Vim were cloned into pSicheck2 vectors. Renilla was used as transfection control. Data were normalized to empty vectors and then to GFP overexpressing ESCs. Shown are mean ± SD, n = 4. P values are as indicated. Unpaired two‐tailed Student's t‐test.
  5. Luciferase assay for 3′UTR of Tgfbr2 with miR‐294 targeting sites mutated. Renilla was used as transfection control. Data were normalized to empty vectors and then to wild‐type 3′UTR reporter in GFP overexpressing ESCs. Shown are mean ± SD, n = 4. P values are as indicated. Unpaired two‐tailed Student's t‐test.
  6. RT–PCR analysis of selected key miR‐294 targets in control or siMbnl1/2 transfected 3T3 cells. The β‐actin gene was used as a control. Data were normalized to the mRNA level of control‐transfected 3T3 cells. Data are presented as mean ± SD, n = 3. P values are as indicated. Unpaired two‐tailed Student's t‐test.