Research Paper Volume 15, Issue 14 pp 7124—7145

Figure 2. LncRNA SPIRE1 knockdown impaired the immunomodulatory properties of normal mandibular BM-MSCs by causing Th17/Treg imbalance. (A–F) Pre-activated CD4⁺ T cells were cultured alone, or co-cultured with mandibular BM-MSCs from control mice or periodontitis mice under the specific Th17 (A–C) or Treg (D–E) polarization condition. Representative flow cytometry profiles of T cells for Th17 (A) or Treg (D) populations identified as CD4⁺IL-17⁺ cells or CD4⁺FoxP3⁺ cells, respectively, are shown, and their percentages among total CD4⁺ cells were summarized. The mRNA levels of RORC (B) or FoxP3 (E) in total T cells and the soluble levels of IL-17 (C) or IL-10 (E) in co-cultured supernatants were quantitated by qPCR or ELISA, respectively. (F) The silence-efficacy of SPIRE1 in normal mandibular BM-MSCs was confirmed by qPCR analysis of cells transfected with negative control (NC)-shRNA or LncRNA SPIRE1-specific shRNA. (G–M) NC-shRNA or SPIRE1-shRNA transfected normal mandibular BM-MSCs were co-cultured with the pre-activated CD4⁺ T cells under the Th17 (G–I) or Treg (J–M) polarization condition. Representative flow cytometry profiles of Th17 (H) and Treg (K) populations, and their percentages are shown. The mRNA levels of RORC (I) or FoxP3 (L), and the levels of IL-17 (I) or IL-10 (M) in supernatants were quantitated by qPCR and ELISA, respectively. n = 3 for each group; ^**P < 0.01; ^***P < 0.001, between the indicated groups.