MLKL Antibody - #DF7412

Fig. 6 The anti-inflammatory effect of Fetuin-A is by inhibiting necroptosis in glutamate-exposed microglial cells. The microglial cells were treated with 100 μm glutamate (Glu) for 24 h to induce cellular injury. Meanwhile, Fetuin-A was treated into medium at indicated concentrations. A, B Expression of RIPK3, MLKL, p-RIPK3 and p-MLKL in microglial cells under various treatment conditions. GAPDH was used as the loading control. And bar graphs show the results of analysis (by band density analysis) of these proteins (n = 3). C–F Immunofluorescence assessment of p-RIPK3 and p-MLKL expression. Scale bar is 50 μm. The relative immunofluorescence intensity of Fetuin-A was detected with Image-J software (n = 3). G RIPK1 was immunoprecipitated with its antibody and resulted in co-immunoprecipitation of RIPK3. Immunoprecipitation of RIPK3 with its antibody caused co-immunoprecipitation of RIPK1 in microglial cells (n = 3). H Transmission electron microscopy (TEM) images of tissues. Translucent cytoplasm, mitochondrial swelling and destruction of membrane integrity were observed in TNF-α + zVAD-treated microglial cells (scale bar = 5 or 1 μm, n = 3). I IL-6, IL-1β, TNF-α, and IL-10 secretion was measured using ELISA (n = 5). Data are presented as the means ± SD; *P 

Figure 7. OTUB1 is involved in cisplatin resistance of BLCA through β-catenin stabilization. A. Cell survival assay was determined in T24 control cell and T24 cisplatin-resistance cell. B. The changes in cell morphology between the T24 control cell and the T24 cisplatin-resistance cell. C. Relative expression of several chemoresistance markers (including MDR1, BCRP and YB-1) in T24 control cell and T24 cisplatin-resistance cell with or without cisplatin treatment. D. Relative expression of OTUB1 in T24 control cell and T24 cisplatin-resistance cell with or without cisplatin treatment. E. Relative expression of OTUB1, necroptosis-related markers (such as RIPK3, MLKL and P-MLKL), β-catenin and downstream proteins (including AXIN-2, C-myc, cyclin D1 and TCF1) in T24 cisplatin-resistance cells treated with gradient cisplatin concentration. F. Immunoprecipitation assay showed the relationship between OTUB1 and β-catenin in T24 cisplatin-resistance cells. G. Immunoprecipitation assay showed that gradient cisplatin concentration promotes the interaction between OTUB1 and β-catenin, and restrains the ubiquitination of β-catenin. H. Relative expression of OTUB1, necroptosis-related markers (such as RIPK3, MLKL and P-MLKL), β-catenin and downstream targets following elevated OTUB1, β-catenin with or without XAV-939 treatment in T24 cisplatin-resistance cell (control, OTUB1, OTUB1/XAV-939, β-catenin). I. Cell survival assay was determined in T24 cisplatin-resistance cells following overexpressed OTUB1 with or without 10uM XAV-939 (or knockdown OTUB1 with or without overexpressed β-catenin). J. Knockdown OTUB1 restrains the growth in T24 cisplatin-resistance mice bladder tumor in vivo.

Fig. 8 |Changes of RIP1, RIP3 and MLKL at mRNA and protein levels in the different groups (means ± SD, n=6) A, B, C: RIP1, RIP3 and MLKL mRNA levels in cardiomyocytes normalized by GAPDH levels. **P<0.01 compared with NG; #P<0.05, ##P<0.01 compared with HG; ^P<0.05, ^^P<0.01 compared with HG+Alda-1 D: Representative western blots of RIP1, RIP3 and MLKL proteins