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: SNHG16 deletion impairs cell cisplatin resistance and malignant phenotypes by sponging miR-338-3p in neuroblastoma. SK-N-AS-R and SK-N-SH-R cells were transfected with si-NC, si-SNHG16, si-SNHG16 + in-miR-NC, si-SNHG16 + in-miR-338-3p. a The relative expression of miR-338-3p was measured using qRT-PCR. b The IC50 value for cisplatin was assessed by CCK-8 assay. c Western blot assay was used to detect the expression of drug-resistance associated proteins MRP1 and P-gp. d, e Cell proliferation was examined using CCK-8 assay. f, g Transwell assay was used for the detection of cell migration and invasion ability. h Cell apoptosis was analyzed using Flow cytometric analysis. Each experiments were repeated three times independently, and the average was taken. *P < 0.05
Fig 3: SNHG16 is up-regulated and miR-338-3p is down-regulated in cisplatin resistant neuroblastoma tissues and cells. a, b The expression of SNHG16 and miR-338-3p was detected using qRT-PCR in cisplatin resistant and sensitive neuroblastoma tumor tissue. c, d The IC50 value for cisplatin was calculated by CCK-8 assay in SK-N-AS-R and SK-N-SH-R cells. e Western blot analysis of the levels of MRP1 and p-gp protein in cisplatin-resistant neuroblastoma cell lines SK-N-AS-R and SK-N-SH-R and parental SK-N-AS and SK-N-SH was performed. f, g The expression of SNHG16 and miR-338-3p was detected using qRT-PCR in cisplatin-resistant neuroblastoma cell lines (SK-N-AS-R and SK-N-SH-R) and corresponding parental neuroblastoma cell lines (SK-N-AS and SK-N-SH). The same experiment was repeated three times, and the average was taken. *P < 0.05
Fig 4: SNHG16 deletion inhibits cell cisplatin resistance and malignant phenotypes in neuroblastoma. SNHG16 was silenced in SK-N-AS-R and SK-N-SH-R cells using siRNA sequences targeting SNHG16. a The relative expression of SNHG16 was measured using qRT-PCR. b The IC50 value for cisplatin was assessed by CCK-8 assay. c The expressions of drug-resistance associated proteins MRP1 and P-gp were examined by western blot assay. d The CCK-8 assay was performed to detect cell proliferation. e, f Transwell assay was used to determine cell migration and invasion ability. g Cell apoptosis was analyzed using Flow cytometric analysis. The same experiment was repeated three times, and data represented as the average of three independent replicates. *P < 0.05
Fig 5: PLK4 overexpression reverses miR-338-3p re-expression-mediated inhibition on cisplatin resistance and tumorigenesis in neuroblastoma cells. SK-N-AS-R and SK-N-SH-R cells were transfected with miR-NC, miR-338-3p, miR-338-3p + pcDNA, or miR-338-3p + PLK4. a Western blot was used to detect the expression of PLK4. b The IC50 value for cisplatin was calculated by CCK-8 assay. c The expressions of drug-resistance associated proteins MRP1 and P-gp were examined by western blot assay. d, e The CCK-8 assay was performed to detect cell proliferation. f, g Transwell assay was used to determine cell migration and invasion ability. h The apoptosis rate was analyzed using Flow Cytometry. Each experiment was performed for three times, and results represented as the average of three independent replicates. *P < 0.05
Supplier Page from Abcam for Anti-MRP1 antibody [EPR21062]