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Fig. 4 | Stem Cell Research & Therapy

Fig. 4

From: DEPTOR regulates osteogenic differentiation via inhibiting MEG3-mediated activation of BMP4 signaling and is involved in osteoporosis

Fig. 4

Knockdown of DEPTOR elevates MEG3 expression to regulate osteogenesis. a Hierarchical clustering indicated MEG3 was differentially expressed (P < 0.05) in DEPTOR knockdown hBMSCs. b Relative MEG3 expression measured by qRT-PCR in DEPTOR knockdown hBMSCs. GAPDH used for normalization. c–f MEG3 siRNA (si-MEG3) transfected into hBMSCs with DEPTOR knockdown. NC used as negative control. c Relative MEG3 expression determined by qRT-PCR. GAPDH used for normalization. d ALP staining on day 7 and ARS staining on day 14 after osteogenic induction. e, f ALP activity on day 7 and AZR mineralization assay on day 14 after osteogenic induction. Data presented as mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001 (n = 3 independent experiments). See Additional file 4: Figure S3. ALP alkaline phosphatase, ARS/AZR Alizarin red S, DEPTOR DEP domain containing mTOR interacting protein, MEG3 maternally expressed 3 (nonprotein coding), shDEPTOR #1/shDEPTOR #2 lentivirus expressing shRNAs targeting DEPTOR, shNC scrambled control, sh short hairpin, si short interfering

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