Fig 1: Schematic diagram of the mechanism by which the circ_0006168/miR-194-5p/JMJD1C regulates cell proliferation, migration, invasion, and Taxol resistance in Taxol-resistant ESCC cells
Fig 2: Circ_0006168 knockdown repressed tumor growth in vivo. Eca109/Taxol and KYSE150/Taxol cells transfected with sh-control or sh-circ_0006168 were injected into nude mice to establish mice xenograft model. A, B Tumor volume and weight were measured. C, D The expression levels of circ_0006168 and miR-194-5p were measured by qRT-PCR in the collected tissues. E Western blot assay was used to analyze the protein expression of JMJD1C in the collected tissues. **P < 0.001, ***P < 0.0001
Fig 3: Circ_0006168 regulated JMJD1C expression by acting as a sponge of miR-194-5p. A Predicted binding sites between miR-194-5p and circ_0006168 were shown. B The expression of miR-194-5p was analyzed by qRT-PCR in Eca109/Taxol and KYSE150/Taxol transfected with si-control and si-circ_0006168. C The level of miR-194-5p was examined by qRT-PCR in Eca109/Taxol and KYSE150/Taxol cells transfected with miR-NC or miR-194-5p. D Dual-luciferase reporter assay was performed to determine the luciferase activity in Eca109/Taxol and KYSE150/Taxol cells co-transfected with miR-NC or miR-194-5p and circ_0006168-wt or circ_0006168-mut. E Circ_0006168 and miR-194-5p enrichment in Eca109/Taxol and KYSE150/Taxol were analyzed by RIP assay. F Circ_0006168 enrichment in Eca109/Taxol and KYSE150/Taxol transfected with NC-input, bio-NC, bio-miR-194-5p, or miR-194-5p-input was detected using RNA pull-down assay. G The level of miR-194-5p was detected by qRT-PCR in ESCC tissues and adjacent normal tissues. H The expression of miR-194-5p was examined by qRT-PCR in HET-1A cells, parental cells (Eca109 and KYSE150), and Taxol-resistant cells (Eca109/Taxol and KYSE150/Taxol). I The putative binding sites between miR-194-5p and JMJD1C were predicted by TargetScan. J The luciferase activity was determined in Eca109/Taxol and KYSE150/Taxol co-transfected with miR-NC or miR-194-5p and JMJD1C-wt or JMJD1C-mut. K JMJD1C and miR-194-5p enrichment in Eca109/Taxol and KYSE150/Taxol were measured by RIP assay. L Circ_0006168 enrichment was determined by RNA pull-down assay in Eca109/Taxol and KYSE150/Taxol cells transfected with NC-input, bio-NC, bio-miR-194-5p, or miR-194-5p-input. M The expression of miR-194-5p was examined by qRT-PCR Eca109/Taxol and KYSE150/Taxol cells transfected with inhibitor-NC or inhibitor-miR-194-5p. N The protein expression of JMJD1C was tested by western blot analysis in Eca109/Taxol and KYSE150/Taxol transfected with miR-NC, miR-194-5p, inhibitor-NC, or inhibitor-miR-194-5p. O Western blot assay was carried out to analyze the protein expression of JMJD1C in Eca109/Taxol and KYSE150/Taxol transfected with si-control, si-circ_0006168, si-circ_0006168 + inhibitor-NC, si-circ_0006168 + inhibitor-miR-194-5p. *P < 0.05, **P < 0.001, ***P < 0.0001
Fig 4: Circ_0006168 regulated Taxol resistance by affecting JMJD1C expression. Eca109/Taxol and KYSE150/Taxol cells were transfected with si-control, si-circ_0006168, si-circ_0006168 + pcDNA, or si-circ_0006168 + pcDNA-JMJD1C. A Western blot assay was used to analyze the protein abundance of JMJD1C. B, C Cell viability and IC50 value of Taxol were determined by CCK-8 analysis. D, E CCK-8 assay and colony formation assay were employed to evaluate cell proliferation. F, G Cell migration and invasion abilities were assessed by transwell assay. H Flow cytometry analysis was applied to detect cell apoptosis. *P < 0.05, **P < 0.001, ***P < 0.0001
Fig 5: JMJD1C silence improved Taxol sensitivity in Taxol-resistant cells. A The mRNA expression of JMJD1C was analyzed by qRT-PCR in ESCC tissues and adjacent normal tissues, and western blot assay was performed to measure the protein expression of JMJD1C in HET-1A cells, parental cells (Eca109 and KYSE150), and Taxol-resistant cells (Eca109/Taxol and KYSE150/Taxol). B–I Eca109/Taxol and KYSE150/Taxol cells were transfected with si-control or si-JMJD1C. B The protein expression of JMJD1C was detected by western blot assay. C, D CCK-8 assay was employed to detect cell viability and IC50 value of Taxol. E Cell viability was measured using CCK-8 assay. F Colony formation assay was utilized to assess cell colony formation ability. G, H The migration and invasion capacities were evaluated using transwell assay (× 100). I Flow cytometry analysis was utilized to determine cell apoptosis. *P < 0.05, **P < 0.001, ***P < 0.0001
Supplier Page from Abcam for Anti-JMJD1C antibody