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. 2020 Aug 27;29:13–22. doi: 10.1016/j.jare.2020.08.013

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© 2020 The Authors. Published by Elsevier B.V. on behalf of Cairo University.

This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

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Fig. 4 — HRC^+/+ attenuated the effects of vitamin D on cell migration, proliferation and apoptosis. (A) Western blots show HRC overexpression in H460 cells; a 200-fold increase in HRC expression was noted. (B) Overexpressed HRC detected by real time RT-PCR. (C) The histogram shows HRC protein expression after 24 h incubation with vitamin D. (D) Effects of vitamin D on HRC mRNA expression determined by real time RT-PCR. (E) Wound healing assay. Vitamin D with HRC^+/+ partially reduced cell mobility compared with that with HRC^+/+, but the effect was weaker than that in the VD group. (F) Transwell migration assays. The histogram shows that vitamin D with HRC^+/+ partially inhibited H460 cell migration. (G) Assays of H460 cell proliferation showed that HRC^+/+ attenuates the effects of vitamin D on cell migration and proliferation. (H) Apoptosis induction assessed by flow cytometry after staining with annexin V-PE/7-AAD. Vitamin D-induced H460 cell apoptosis was significantly reduced by HRC^+/+. Values are expressed as means ± SD. Statistical significance is shown as *P < 0.05, **P < 0.01, and ***P < 0.001. HRC, Histidine-rich calcium binding protein.