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. 2022 Aug 8;8:353. doi: 10.1038/s41420-022-01146-7

Fig. 1. Cpd16 activates Nrf2 signaling cascade in osteoblasts.

Fig. 1

Primary murine osteoblasts (A–D, G–I), human osteoblasts (J–O), or the MC3T3-E1 murine osteoblastic cells (P, Q) were stimulated with Cpd16, ARE activity and NQO1 activity as well as cell viability were measured (A); Keap1-Nrf2 association was measured through co-immunoprecipitation (Co-IP) (B, J); Proteins in cytosol/nuclear fraction lysates were examined (C, G, I, K, M, O, P), with mRNAs measured by qRT-PCR (D, H, L, N, Q). The primary murine osteoblasts were treated with MG-132 (10 μM) or plus Cpd16 (25 μM) for 8 h, total protein lysates were tested (E). The murine osteoblasts were pretreated for 1 h with cycloheximide (CHX, 25/100 μg/mL), following by Cpd16 (25 μM) stimulation for another 8 h, listed proteins were shown (F). “C” is untreated control (same for all Figures). “Veh” is vehicle control (0.1% DMSO) (same for all Figures). *P < 0.05 versus “Veh” cells.