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Table S1 PCR primers for human adipogenic and osteogenic specific genes

Table S2 PCR primers for human genes of Ca2+ signals and transporters

Table S3 PCR primers for human specific genes related to cADP ribose

Table S4 siRNA sequences of TRPM2 (NM_003307)

Table S5 Flow cytometry mark results -- human bone marrow MSCs

Fig. S1 Ca2+ signaling pathways in human MSCs. (A) The IP3Rs blocker 2-APB suppressed spontaneous Ca2+i oscillations (n = 27). (B) SERCAs inhibitor CPA abolished Ca2+i oscillations (n = 27). (C) RyRs blocker showed no significant effect on Ca2+i oscillations (n = 24). (D) LaCl3, a SOC entry blocker blocked Ca2+i oscillations (n = 25). (E) RT-PCR results show significant mRNAs for IP3R1-3, SERCA1-3, PMCA1-3, NCX1, NCX3 and NCX4, but not for RyRs. (F) Messenger RNAs for RyR1-3 were detectable human SH-SY5Y neuroblastoma cells (H), but not in human MSCs (S) using three primers of RyRs in (E). IP3R: IP3 receptor; SERCA: sarco/endoplasmic reticulum Ca2+-ATPase; PMCA: plasma membrane Ca2+-ATPase. NCX: Na+-Ca2+ exchanger; RyR: ryanodine receptor; H: total RNA extracted from human SH-SY5Y neuroblastoma cells; S: human MSCs.

Fig. S2 Effects of cADP ribose on adipogenesis in human MSCs. (A) Images showing adipogenic differentiation of human MSCs at induction cycle 1, 2 and 3. cADP ribose showed no significant effect on adipogenesis in human MSCs. Similar results were obtained in a total of three experiments. (B) No difference was found in PPAR-γ protein expression in the differentiated adipocytes (day 9) with and without the treatment of 50 μM cADPR (C) No spontaneous Ca2+i oscillations were observed during adipogenesis on day 1, day 4 and day 9. cADP ribose did not initiate Ca2+i transient or Ca2+i oscillations in these cells (n = 32 for each).

Fig. S3 Effects of cADP ribose on osteogenesis in human MSCs. (A) Images showing the osteogenesis on day 10, day 15 and day 20 by Alizarin red S staining. cADP ribose had no significant effect on osteogenesis in human MSCs. Similar results were obtained in a total of three experiments. (B) No difference was found in osteocalcin protein expression in the differentiated osteocytes (day 18) with and without the treatment of 50 μM cADPR (C) No spontaneous Ca2+i oscillations were observed during osteogenesis on day 1, day 9 and day 18. Cyclic ADP ribose did not initiate Ca2+i transient or Ca2+i oscillations in these cells (n = 32 for each).

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