P Glycoprotein 1（MDR1） Antibody - #AF5185

Figure 4. Cytotoxicity of different drug formulations. (a) MTT assay of HepG2 cells and the cocultured model. (b) Cell viability of HepG2 cells. (c) Cell viability of the cocultured system. Scale bars represent 100 μm. (d) Live/dead staining of HepG2 cells, where green- and red-dyed cells represent live and dead HepG2 cells, respectively. (e) Live/dead staining of the cocultured model, where the yellow-, red-, and green-colored cells denote the dead LX-2, dead HepG2, and live LX-2 cells, respectively. Scale bars represent 100 μm. (f) Western blotting assay for the expression of P-gp. (g) Quantitative analysis of P-gp. *P < 0.05, **P < 0.01, ***P < 0.001.

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. 2| Autophagy is increased and apoptosis is reduced in the EOC re-ascites group. a IHC analysis of LC-3, Belin-1, CASP-9, and c-CASP-3 expression in EOC ascites cells (scale bar =30µm). b, c Western blot analysis of autophagy/apoptosis-related proteins expressed in the chemosensitive group and re-ascites group (***p<0.001, **p<0.01, *p<0.05). d Immunofluorescence analysis of the chemosensitive group and re-ascites group with LC3 (green) and LAMP2 (red) staining. Nuclei were counterstained with DAPI (blue). e qRT-PCR analysis of the average relative mRNA expression levels of autophagy/apoptosis-related genes in the chemosensitive group and re-ascites group (**p<0.01, *p<0.05)

Figure 6 Western blotting against α-SMA, Bcl-2, and P-gp in total soluble protein fractions from (1) control 4T1 cells in monoculture, (2) control co-cultures of 4T1 cells and NIH 3T3 fibroblasts, or co-cultures treated with (3) free CEL, (4) free CEL+BA, (5) CL, (6) CL@BM or (7) F/CL@BM. (A) Western blotting assay for the expression of protein. (B) Quantitative analysis. ***p < 0.001, ****p < 0.0001.

FIGURE 3  miR-506 down-regulated MDR1/P-gp expression in HCT116-OxR. A, The mRNA level of MDR1 was decreased after transfection with the miR-506 mimic of the relative chemoresistant genes as demonstrated by qRT-PCR. B, The protein level of MDR1 was decreased after transfection with the miR-506 mimic of the relative chemoresistant proteins as demonstrated by Western blot. C, Expression of P-gp detected by immunofluorescence staining.HCT116-OxR-miR-506 cells showed low levels of fluorescent staining of P-gp, whereas maximal staining of P-gp was observed in HCT116-OxR cells, readily distinguished from background. Zoom: 200×. *P<.05)