Fig. 5

TNFAIP6 defines a subpopulation of mouse MSCs with enhanced immune suppression activities. A Cell morphology of CD45⁻TNFAIP6⁻ and CD45⁻TNFAIP6⁺ MSCs. B Left panel: mRNA level of TNFAIP6 was assessed by qPCR (n = 3); Right panel: protein level of TNFAIP6 was assessed by western blot. C Protein level of TNFAIP6 secreted from the CD45⁻TNFAIP6⁻ or CD45⁻TNFAIP6⁺ MSCs (n = 3). D Splenocytes proliferation assay after co-culture with CD45⁻TNFAIP6⁻ or CD45⁻TNFAIP6⁺ MSCs (n = 3). Both high cell number (5 × 10⁴) and low cell number (2 × 10⁴) were assessed. E Splenocytes proliferation assay after co-culture with CD45⁻TNFAIP6⁻ or CD45⁻TNFAIP6⁺ MSCs was assessed with CFSE approach. High cell number (5 × 10⁴) was used. F CD44 expression of CD45⁻TNFAIP6⁻ or CD45⁻TNFAIP6⁺ MSCs, determined by flow cytometry. G Cell proliferation of CD45⁻TNFAIP6⁻ or CD45⁻TNFAIP6⁺ MSCs was determined by doubling time (n = 3). H The adipocytes differentiation efficiency was assessed by Oil Red O staining. I The adipocytes differentiation efficiency was quantified by Oil Red O staining and qPCR analysis of gene Lpl and Pparγ (n = 3). J The osteocytes differentiation efficiency was assessed by Alizarin Red staining. K The osteocytes differentiation efficiency was quantified by Alizarin Red staining and qPCR analysis of gene Osterix and Runx2 (n = 3). L The chondrocytes differentiation efficiency was assessed by Alcian Blue staining. M The chondrocytes differentiation efficiency was quantified by Alcian Blue staining and qPCR analysis of gene Sox9 and Bmp2 (n = 3). * indicates P < 0.05. TNFAIP6 tumor necrosis factor alpha-induced protein 6; MSCs mesenchymal stromal/stem cells; Lpl lipoprotein lipase; Pparγ peroxisome proliferator-activated receptor gamma; Osterix Sp7 transcription factor; Runx2 RUNX family transcription factor 2; Sox9 SRY-box transcription factor 9; and Bmp2 bone morphogenetic protein 2