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. 2019 Aug 2;42(4):1343–1354. doi: 10.3892/or.2019.7258

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Copyright: © Tan et al.

This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.

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Figure 3. — TRP14 knockdown suppresses autophagy-induced cisplatin resistance in SKOV3/DDP and A2780/DDP cells. (A) The levels of TRP14 were detected by western blot analysis in A2780, A2780/DDP, SKOV3 and SKOV3/DDP cells. **P<0.01 indicates statistical significance vs. normal A2780 and SKOV3 cells. (B) A2780/DDP and SKOV3/DDP cells were transfected with TRP14 shRNA. **P<0.01 indicates statistical significance vs. TRP14-knockdown cells. (C) Autophagy-associated genes (LC3, Beclin 1 and ATG5) were examined in cells in which TRP14 was knocked down cells by western blot analysis. **P<0.01 indicates statistical significance vs. TRP14-knockdown cells. (D) Knockdown of TRP14 with shRNA2 in SKOV3/DDP and A2780/DDP cells. The cells were then transfected with GFP-RFP-LC3-plasmid for overnight. Representative images of GFP-RFP-LC3-II-positive puncta were obtained using a confocal fluorescence microscope. *P<0.05, statistical significance vs. TRP14-knockdown cells. (E) Autophagosome and autolysosome vesicles of A2780/DDP and SKOV3/DDP cells were visualized by transmission electron microscopy following transfection with TRP14 shRNA. The white arrows indicate the typical images of autophagosomes and autolysosomes. TRP14 knockdown suppresses autophagy-induced cisplatin resistance in SKOV3/DDP and A2780/DDP cells. (F) MDC staining of A2780/DDP and SKOV3/DDP cells following the knockdown of TRP14. (G) SKOV3/DDP cells were transfected with TRP14 shRNA2. The cells were then treated with or without 200 nM rapamycin (Rapa) for 24 h. After 24 h, the cells were treated with the indicated concentrations of cisplatin (from o to 16 µg/ml) for 24 h and cell viability was measured by CCK-8 assay. The Kruskal-Wallis was used for multiple comparisons and then Mann Whitney U test and Bonferroni's correction were applied. The values are presented as the means ± SEM (n=3). *P<0.05, statistical significance vs. TRP14-knockdown cells. (H) A2780/DDP cells were transfected with TRP14 shRNA2 overnight. The cells were then treated with or without 200 nM Rapa for 24 h. After 24 h, the cells were treated with the indicated concentrations of cisplatin (from 0 to 16 µg/ml) for 24 h and cell viability was measured by CCK-8 assay. The Kruskal-Wallis was used for multiple comparisons and then Mann Whitney U test and Bonferroni's correction were applied. The values are presented as the means ± SEM (n=3). *P<0.05, statistical significance vs. TRP14-knockdown cells.

Figure 3. — TRP14 knockdown suppresses autophagy-induced cisplatin resistance in SKOV3/DDP and A2780/DDP cells. (A) The levels of TRP14 were detected by western blot analysis in A2780, A2780/DDP, SKOV3 and SKOV3/DDP cells. **P<0.01 indicates statistical significance vs. normal A2780 and SKOV3 cells. (B) A2780/DDP and SKOV3/DDP cells were transfected with TRP14 shRNA. **P<0.01 indicates statistical significance vs. TRP14-knockdown cells. (C) Autophagy-associated genes (LC3, Beclin 1 and ATG5) were examined in cells in which TRP14 was knocked down cells by western blot analysis. **P<0.01 indicates statistical significance vs. TRP14-knockdown cells. (D) Knockdown of TRP14 with shRNA2 in SKOV3/DDP and A2780/DDP cells. The cells were then transfected with GFP-RFP-LC3-plasmid for overnight. Representative images of GFP-RFP-LC3-II-positive puncta were obtained using a confocal fluorescence microscope. *P<0.05, statistical significance vs. TRP14-knockdown cells. (E) Autophagosome and autolysosome vesicles of A2780/DDP and SKOV3/DDP cells were visualized by transmission electron microscopy following transfection with TRP14 shRNA. The white arrows indicate the typical images of autophagosomes and autolysosomes. TRP14 knockdown suppresses autophagy-induced cisplatin resistance in SKOV3/DDP and A2780/DDP cells. (F) MDC staining of A2780/DDP and SKOV3/DDP cells following the knockdown of TRP14. (G) SKOV3/DDP cells were transfected with TRP14 shRNA2. The cells were then treated with or without 200 nM rapamycin (Rapa) for 24 h. After 24 h, the cells were treated with the indicated concentrations of cisplatin (from o to 16 µg/ml) for 24 h and cell viability was measured by CCK-8 assay. The Kruskal-Wallis was used for multiple comparisons and then Mann Whitney U test and Bonferroni's correction were applied. The values are presented as the means ± SEM (n=3). *P<0.05, statistical significance vs. TRP14-knockdown cells. (H) A2780/DDP cells were transfected with TRP14 shRNA2 overnight. The cells were then treated with or without 200 nM Rapa for 24 h. After 24 h, the cells were treated with the indicated concentrations of cisplatin (from 0 to 16 µg/ml) for 24 h and cell viability was measured by CCK-8 assay. The Kruskal-Wallis was used for multiple comparisons and then Mann Whitney U test and Bonferroni's correction were applied. The values are presented as the means ± SEM (n=3). *P<0.05, statistical significance vs. TRP14-knockdown cells.