Fig 1: Mindin knockdown inhibited the activation of TLR4/JNK/ NF-κB signaling after HR in vitro. A TLR4, Mindin, phosphorylated JNK, phosphorylated P65, and phosphorylated IκBα measured by western blot analysis in si-Mindin, si-TLR4 and si-JNK cells induced by HR respectively. B–F Quantitative analysis of TLR4, Mindin, phosphorylated JNK, phosphorylated P65, and phosphorylated IκBα protein expression. *P < 0.05, **P < 0.01 versus HR. G Quantitative analysis of TLR4, JNK, P65, TNF-α and MCP-1 mRNA expression in si-Mindin, si-TLR4 and si-JNK cells induced by HR respectively. ***P < 0.001 versus control HR group
Fig 2: Mindin overexpression promoted production of inflammatory mediators and activation of TLR4/JNK/ NF-κB signaling after HR in vitro. A Representative photomicrographs showing the immunofluorescence of GFP in adGFP and adMindin cells. B Mindin protein expression in adGFP and adMindin cells. C–E Protein levels of TNF-α and MCP-1 in the media of cultured TECs by Western blot. *P < 0.05 versus adGFP Con or adMindn Con. #P < 0.05 versus adGFP HR. F TLR4, phosphorylated JNK, phosphorylated P65, and phosphorylated IκBα measured by western blot analysis in adGFP and adMindin cells induced by HR or not. G–J Quantitative analysis of TLR4, phosphorylated JNK, phosphorylated P65, and phosphorylated IκBα protein expression. *P < 0.05 versus adGFP Con or adMindn Con. #P < 0.05 versus adGFP HR. K Quantitative analysis of TLR4, JNK, P65, TNF-α and MCP-1 mRNA expression in mindin-overexpressed HK-2 cells. ***P < 0.001 versus adGFP HR group
Fig 3: Mindin expression was elevated in renal IR injury in vivo and in vitro. A Mindin measured by western blot analysis in sham mice and IR mice. Quantitative analysis of mindn protein expression. Expression levels were normalized to GAPDH and are presented relative to the sham-operated kidney. *P < 0.05 versus Sham. B Representative photomicrographs showing IHC analysis for mindin after 30 min renal ischemia and 24 h reperfusion (magnification 400 ×). C Mindin measured by western blot analysis in HK-2 cells after 24 h hypoxia and 2, 4, and 8 h reoxygenation. Quantitative analysis of mindin protein expression. Expression levels were normalized to GAPDH and are presented relative to the control cells. *P < 0.05 versus Control. D Representative photomicrographs showing immunofluorescence staining for mindin after 24 h hypoxia and 8 h reoxygenation (magnification 400 ×)
Fig 4: Mindin deficiency limited renal IR induced inflammatory cell infiltration and proinflammatory mediators production. A Representative photomicrographs showing IHC staining of LY6G in WT and mindin KO mice induced by IR or not (magnification 400 ×). B Quantitative analysis of LY6G in the indicated groups. *P < 0.05 versus WT Sham or KO Sham. #P < 0.05 versus WT IR. C Representative photomicrographs showing the IHC staining of CD68 in WT and mindin KO mice induced by IR or not (magnification 400 ×). D Quantitative analysis of CD68 in the indicated groups.*P < 0.05 versus WT Sham or KO Sham. #P < 0.05 versus WT IR. E Representative photomicrographs showing IHC staining of TNF-α in WT and mindin KO mice induced by IR or not (magnification 400 ×). F Quantitative analysis of TNF-α in the indicated groups.*P < 0.05 versus WT Sham or KO Sham. #P < 0.05 versus WT IR. G Representative photomicrographs showing IHC staining of MCP-1 in WT and mindin KO mice induced by IR or not (magnification 400 ×). H Quantitative analysis of MCP-1 in the indicated groups. *P < 0.05 versus WT Sham or KO Sham. #P < 0.05 versus WT IR. I, J Quantitation of TNF-α and MCP-1 in serum was measured by ELISA. *P < 0.05 versus WT Sham or KO Sham. #P < 0.05 versus WT IR
Fig 5: Mindin inhibited cell proliferation significantly. The effect of Mindin overexpression or knockdown on cell proliferation was studied in HK-2 cell. O-NC: adGFP normal control group, si-NC: siRNA normal control group, O-Mindin: Mindin overexpression group, si-Mindin: Mindin siRNA knockdown group. *P < 0.05, **P < 0.01 versus si-NC group; #P < 0.05, ##P < 0.01 versus O-NC group. Interaction between the time and gene interference factors, F = 49.448, P < 0.001; main effect of the time factor, F = 1298.047, P < 0.001
Supplier Page from MedChemExpress for Spondin-2/SPON2 Protein, Human (HEK293, His)