Figure 5
XRCC recruitment rate and ligation activity during the osteogenesis of hMSCs. Hsp70, LIG3, and PARP1 interact with the scaffold protein XRCC1. (A) The protein–protein interaction among Hsp70, LIG3, PARP1, and XRCC was evaluated via co-immunoprecipitation. Full-length blots/gels are presented in Supplementary Fig. 4. (B) Relative interaction levels with XRCC1 are quantified and plotted into a graph. (C) LIG3 and HSP70 levels were analyzed using cell extracts from hMSCs or differentiated cells and (D) plotted into a graph. Full-length blots/gels are presented in Supplementary Fig. 5. (E) Schematic representation of the ligation activity assay. (F) LIG3-activity assay was performed using cell extracts from hMSCs, p. Os, and m. Os. The relevant time points are indicated above each sample. Each blot was electrophoresed using mini-gels. (G) Each activity was quantified and plotted into a graph. T4 Lig is the positive control that contains T4 ligase from Enterobacteria phage T4, and [-] represents the negative control. hMSC, proliferating hMSC; Pre.O, pre-osteoblast; M.Os, mature osteoblast S, Substrate; P, Product. *Represents significant differences as compared with the MSC group.
