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. 2022 Oct 26;13:6345. doi: 10.1038/s41467-022-33946-x

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© The Author(s) 2022

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 7 — a Drug response associated with the autophagy status in SKCM. The orange point indicates sensitivity; the red point indicates resistance. b, d Dose-response curves for the mean value of cell viability of etoposide and BMS536924 in rapamycin-induced (b) or starvation-induced (d) and non-induced conditions in the melanoma cell line A375 and SK-MEL-28. Cell viability was normalized to the level of cells treated with DMSO (b) or DMEM (d). DMEM: Dulbecco’s Modified Eagle Medium for control. EBSS: Earle’s Balanced Salt Solution for starvation. Error bars indicate the mean ± SD. The drug screen data of different groups (n = 4) were fitted and compared by sigmoidal dose-response curves. c Western blot of autophagy markers in A375 and SK-MEL-28 cell lines treated with DMSO, rapamycin, etoposide, and rapamycin + etoposide. The ratios of the LC3-II/LC3-I and p62 band intensities normalized to DMSO are displayed below the blots. Similar results were observed in at least three independent experiments. e The design of xenograft experiments. Nude mice were injected with 2 × 10⁶ A375 cells. When the tumor size reached 50–100 mm³, mice were treated with vehicle, 0.75 mg/kg rapamycin intraperitoneally (i.p.), 5 mg/kg etoposide (i.p.), or the combination of rapamycin and etoposide every 3 days (n = 7). Mice were depicted using “mouse-animal-rodent-mammal-little” (https://pixabay.com/zh/vectors/mouse-animal-rodent-mammal-little-310756/), available under the Simplified Pixabay License (https://pixabay.com/zh/service/license/). f Excised tumors to represent the tumor size on day 18. g Western blot of autophagy markers in mouse tissues with the treatment of vehicle, rapamycin, etoposide, and rapamycin + etoposide. The ratios of the LC3-II/LC3-I and p62 band intensities normalized to vehicle are displayed below the blots (n = 3). h Quantitative analysis of tumor volume at different time points with the treatment (n = 7). Error bars indicate the mean ± SD. The difference in multiple groups was estimated by one-way ANOVA analysis. P values are as follows: p = 0.011 (Vehicle vs. Etoposide), p = 0.0012 (Vehicle vs. Rapamycin), p = 0.026 (Etoposide vs. Rapamycin+Etoposide), and p = 0.00058 (Rapamycin vs. Rapamycin+Etoposide). c, g See Supplementary Fig. 8 for uncropped data.