Fig 1: Osteoblastogenic effects of PDGFB. A Protein expression of GRB2 in FOB1.19 cells and ADSCs. Western blotting was performed under the same conditions (equal exposure time and dose). B Top pathway enrichment in the KEGG database in FOB1.19 cells. C Collagen expression of FOB1.19 in the two groups (PDGFB − , PDGFB +). D Relative protein level of collagen expression in FOB1.19 in the two groups (PDGFB − , PDGFB +). E, F Western blot of the collagen expression and RUNX2 with/without the treatments of PDGFB and si-RUNX2. G Possible mechanism of PDGFB pathogenesis in AS. The relative protein levels were compared by unpaired sample t-tests. * P < 0.05
Fig 2: Effects of PDGFB on ADSCs. A The mRNA expression of several osteogenic-related genes in the two groups. B ALP staining of ADSCs in the two groups. C, D ARS staining of ADSCs in the two groups. E Western blot of ADSCs in the two groups. F Relative protein levels of ALP and RUNX2 in ADSCs. G Proliferation in the two groups. H Slope of proliferation in the two groups. I Chemotaxis in the two groups. J Migration results for the two groups. The gene expression levels and protein levels were compared by unpaired sample t-tests. ** P < 0.01, * P < 0.05
Fig 3: Immunochemistry analysis of the spinal entheseal tissues of AS patients and traumatic injury patients. A Elevated expression of the PDGFB, GRB2, P-ERK, and RUNX2 in AS patients. B Relative positive area of target protein in the sections were quantified
Fig 4: Cytokines detected in AS patients and healthy controls. A Top pathway enrichment according to the InterPro database. B Differential expression of six cytokines in AS and healthy controls in validation I stage. C Differential expression of PDGFB in Chinese AS patients and healthy controls in the validation II stage. AS-1 represents the 40 AS patients at baseline in the validation II stage, and AS-2 represents the 60 late-stage AS patients who have had a disease duration longer than 5 years. The expression levels of cytokines were compared by unpaired sample t-tests. *** P < 0.005, ** P < 0.01
Fig 5: Correlation of PDGFB with clinical indices. A Correlation between PDGFB and BASFI. B Correlation between PDGFB and all clinical information. C Correlation between PDGFB and ΔmSASSS in Chinese AS patients in validation II-40. D Expression level of PDGFB secreted from monocytes after stimulation with M-CSF and RANKL for 7 days, which were analyzed by ELISA. E TRAP staining of the osteoclastogenesis assay for 7 days stimulation. The expression levels of PDGFB in the AS group and control group were compared by unpaired sample t-tests. * P < 0.05
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