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. 2021 Oct 24;30:09636897211052959. doi: 10.1177/09636897211052959

Figure 2.

Figure 2.

Knockdown of circRFWD2 inhibited osteogenic differentiation of hDPSCs. (A) Relative expression of circRFWD2 after osteogenic induction for 0, 7, and 14 days. (B) qRT-PCR was performed to determine the expression of circRFWD2 in si-NC and si-circRFWD2 groups. (C) ALP staining and ARS staining in si-NC and si-circRFWD2 groups. (D) ALP activity was measured by spectrophotometry and normalized to the si-NC group. (E) The quantity of ARS staining was detected by densitometry at 562 nm. (F) Relative expression of RUNX2 mRNA in si-NC and si-circRFWD2 groups. (G) Relative expression of OPN mRNA in si-NC and si-circRFWD2 groups. (H) Western blot analysis of RUNX2 in si-NC and si-circRFWD2 groups. (I) Quantity of western blot analysis of RUNX2 in si-NC and si-circRFWD2 groups. GAPDH is a protein-loading control. All experiments were repeated at least three times. The data are shown as the mean ± SD (n = 3). *P < 0.05, **P < 0.01. ALP: Alkaline phosphatase; ARS: Alizarin Red S; si-circRFWD2: small interfering RNAs targeting circRFWD2; si-NC: small interfering RNA negative control.