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. 2020 Aug 24;11:364. doi: 10.1186/s13287-020-01877-3

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© The Author(s) 2020

Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 2 — CircSIPA1L1 stimulates DPSC osteogenesis. a Western blot results revealed that the protein levels of OSX, RUNX2, and ALP significantly increased in the circSIPA1L1 group compared with the NC group. b Western blot assay showed higher protein levels of ALP, RUNX2, and OSX in the Si-NC group than the Si-circSIPA1L1-3 group and Si-circSIPA1L1-1 group, respectively. GAPDH was the internal control. c Images of alkaline phosphatase (ALP) staining in the different groups, and Si-NC treatment led to the highest ALP activity. Cells were cultured for 7 days. After 14 days of co-culture, the formation of mineralized nodules in DPSCs in the circSIPA1L1 group generated more calcified nodules than the NC group; meanwhile, the Si-NC group has more calcified nodules than Si-circSIPA1L1-3 group and Si-circSIPA1L1-1 group. d Immunofluorescence staining showed positive expressions of ALP and OSX in DPSCs transfected with si-NC, si-circSIPA1L1-1 or si-circSIPA1L1-3, respectively. *P < 0.05, **P < 0.01. e The mRNA levels of OSX, RUNX2, and ALP in DPSCs measured by RT-PCR following the 3-day osteogenesis. The results showed higher levels of ALP, RUNX2, and OSX in the circSIPA1L1 group than the NC group. f Histograms showed quantification of Alizarin red staining by spectrophotometry of the NC group and circSIPA1L1 group. g H&E staining and Masson staining Si-NC, si-circSIPA1L1-1, and si-circSIPA1L1-3 groups. H&E and Masson staining showed less bone-like structures and collagen deposits in DPSCs of the circSIPA1L1-downexpressing group than the control group. Bone/dentin-like tissues (arrow), S around the scaffold, scale bar = 100 μm. h The results of RT-PCR showed that the expression of ALP, RUNX2, and OSX were decreased in the Si-circSIPA1L1-3 group and Si-circSIPA1L1-1 group compared with the Si-NC group. n = 3. **2^-ΔΔCt > 2, P < 0.01; *1 < 2^-ΔΔCt < 2, P < 0.05. i The quantification of Alizarin red staining by spectrophotometry revealed that si-circSIPA1L1-1 or si-circSIPA1L1-3 showed less calcified nodules than si-NC group