Fig 1: Circ_0001721 knockdown impeded DXR resistance and tumor progression in DXR-resistant OS cells. A KHOS/DXR and MG63/DXR cells were introduced with si-NC, si-circ#1, si-circ#2 or si-circ#3, and the knockdown efficiency was examined by qRT-PCR. B After DXR treatment with gradient concentration, IC50 value was evaluated using CCK-8 assay. C, D CCK-8 assay was used to assess cell viability. E, F Transwell assay was utilized to evaluate cell migration and invasion. G Flow cytometry was used to monitor the apoptosis rate. H The levels of multidrug resistance-related proteins (MRP1, P-gp and LRP) were examined by western blot. I The activity of Wnt/ß-catenin pathway was evaluated by detecting the protein levels of ß-catenin, cyclin D1 and c-myc. J, K The protein levels of MRP1, P-gp, LRP, ß-catenin, cyclin D1 and c-myc were measured in KHOS and MG63 cells transfected with si-NC or si-circ#2. *P < 0.05
Fig 2: Knockdown of CASC9 reverses chemoresistance in BGC823/DR and SGC7901/DR cells(A) IC50 of BGC823/DR and SGC7901/DR cells to paclitaxel after CASC9 knockdown. (B) IC50 of BGC823/DR and SGC7901/DR cells to adriamycin after CASC9 knockdown. (C) The expression of MDR1 protein in BGC823/DR and SGC7901/DR cells after CASC9 knockdown.
Fig 3: Expression of ABCB1 within different human liver regions. (a) Diagram of the different regions where samples taken within the same liver; (b) The mRNA expression of ABCB1 was determined within different regions of 14 different livers; (c) Protein expression of ABCB1 within different human liver regions: (distal large lobe n = 8; distal small lobe n = 6), medial large lobe n = 7; medial small lobe n = 6, central large lobe n = 10, central small lobe n = 5, APC = adjacent to portal circulation n = 6 and ABD = adjacent to bile duct N = 11); (d) Correlation between mRNA expression and protein expression for ABCB1, with 95 % confidence intervals (dotted line), and data analyzed by Spearman’s correlation; (e) ABCB1 mRNA expression (line = means) within the livers of 10 individuals; (f) The ABCB1 protein expression showing intra-individual variability of protein expression (8 regions for A, D, E, J; 7 regions for B and C; 4 regions for I and 2 regions for F, G and H).
Fig 4: The MDM2/MDMX inhibitor in combination with DOX reduces rhodamine 123 efflux in drug-resistant breast cancer cells. The cells were treated with the indicated agents for 24 h, and intracellular rhodamine 123 levels were measured to explore the transporter activity of the membrane pump protein P-gp. The concentration levels of each agent were used as follows: DOX 0.08 µg/ml in MCF-7/DOX and 0.20 µg/ml in ZR-75-30/DOX, MDM2/MDMX inhibitor 9.58 µg/ml in MCF-7/DOX and 13.70 µg/ml in ZR-75-30/DOX. The representative charts (A) and quantified data (B, C) of the rhodamine 123 assay are shown. The expression levels of the proteins investigated were examined by western blot analysis, and GAPDH was used as a loading control. The representative charts (D) and quantified data (E, F) of the western blot analysis are also shown. The values presented are indicative of the mean ± SD for each group. *P<0.05, **P<0.01, ***P<0.001 (n = 3) vs. the control group, the DOX group and/or the MDM2/MDMX inhibitor group. Immunofluorescence staining was also used in drug-resistant breast cancer cells to confirm the expression of the proteins investigated. The representative charts (G) of three independent experiments are shown.
Fig 5: Hesperetin combined with the nuclear factor-?B signaling pathway inhibitor JSH-23 significantly enhanced the sensitivity of A549/DDP cells to DDP. (A) Compared with cells treated with hesperetin or JSH-23 alone, co-treatment with hesperetin and JSH-23 significantly improved the effect of DDP on A549/DDP cells. (B) IC50 values of DDP on cells treated with hesperetin alone or in combination with JSH-23. IC50 values were significantly reduced when the treatments were combined. (C) Representative blots and (D) densitometry analysis of P-gp protein expression when A549/DDP cells were treated with hesperetin alone or combined with JSH-23. (E) Representative dot plots of apoptosis when A549/DDP cells were treated with hesperetin alone or combined with JSH-23. (F) Quantification of cell apoptosis. *P<0.05 vs. control group. #P<0.05 vs. hesperetin or JSH-23. Error bars indicate standard deviations. n=3. H, hesperetin; DDP, cisplatin; IC50, half-maximal inhibitory concentration; OD, optical density; AVG, Annexin V; PI, propidium iodide.
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