Fig. 1

Schematic overview of the quantitative TMT proteomics workflow and validation of osteogenic differentiation of hPDLSCs. a Experimental overview of proteomic analysis of hPDLSCs during osteogenesis. hPDLSCs were isolated with limiting dilution methods and induced to undergo osteogenic differentiation. Protein lysates of cells induced for 0 (uninduced), 3, 7 and 14 days were collected. Following trypsin digestion of equal amounts of protein, peptides were labelled with 6-plex TMT reagents, fractionated by HPLC and analysed by LC-MS/MS. After protein identification and quantitation, bioinformatic analysis was performed. We performed three biological replicates of each time point. b Morphological characterizations of osteogenic-differentiated hPDLSCs. hPDLSCs induced for 0 (uninduced), 3, 7 and 14 days were imaged in bright field before ALP and ARS staining, respectively. Scale bar 500 μm. c Validation of osteogenesis-related proteins (RUNX2 and ALP) by western blotting. The data are represented as the means ± standard deviation (SD). **p < 0.01. All data were collected at least in triplicate