Journal: Frontiers in Immunology

Article Title: The role of WTAP in regulating macrophage-mediated osteoimmune responses and tissue regeneration in periodontitis

doi: 10.3389/fimmu.2024.1423378

Figure Lengend Snippet: Reducing of WTAP expression in macrophages promotes BMSC osteogenic differentiation. (A) Schematic diagram of the indirect co-culture system between BMDMs and BMSCs. (B-D) RT-PCR analysis quantifying the mRNA levels of osteogenic markers runx2 , bmp2 , ocn , opn and opg in BMSCs co-cultured with conditional medium at 4 (B) , 7 (C) and 14 (D) days (n = 3). (E) Representative images of ALP staining of BMSCs cultured in conditional medium for 14 days. Scale bar = 500 μm. (F) Quantitative analysis of ALP activity of BMSCs cultured in conditional medium for 14 days (n = 3). (G) Representative images of ARS staining of BMSCs cultured in conditional medium for 21 days. Scale bar = 500 μm. (H) Quantitative analysis of ARS staining in BMSCs (n = 3). Data are presented as the mean ± SD from at least three independent experiments. P values were calculated using two-tailed Student’s t test; * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Macrophage-conditioned medium was prepared using the supernatants from shNC- and shWTAP-treated BMDMs, and mixed with osteogenic induction medium for mouse BMSCs (MUXMX-90021, Cyagen) at a ratio of 1:2.

Techniques: Expressing, Co-Culture Assay, Reverse Transcription Polymerase Chain Reaction, Cell Culture, Staining, Activity Assay, Two Tailed Test