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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. 8 — a The mRNA expression of DDIT4 in A375 and SK-MEL-28 by RNA-seq (n = 3). The boxes show the median ±1 quartile, with whiskers extending to the most extreme data point within 1.5 interquartile range from the box boundaries. The significance (p value) of differentially expressed genes was evaluated with ‘DESeq2’ package. b Relative mRNA expression of DDIT4 in A375 and SK-MEL-28 by RT-PCR (n = 3). c Western blot of DDIT4 in A375 or SK-MEL-28 cells treated with si-NC or DDIT4 siRNAs in combination with etoposide. DDIT4 band intensities normalized to si-NC + etoposide are displayed below the blots. d The cell viability of A375 or SK-MEL-28 cells treated with si-NC or DDIT4 siRNAs in combination with etoposide (n = 4). Pink bars denote the successful knockdown of si-DDIT4 (si-DDIT4-2 and si-DDIT4-3), while gray bar denotes the unsuccessful knockdown of si-DDIT4 (si-DDIT4-1). e Western blot of DDIT4 in A375 or SK-MEL-28 cells transfected with DDIT4-OE lentivirus or vector in combination with etoposide and rapamycin. DDIT4 band intensities normalized to Vector+Etoposide are displayed below the blots. f The cell viability of A375 or SK-MEL-28 cells transfected with DDIT4-OE lentivirus or vector in combination with etoposide and rapamycin (n = 4). g The cell viability of A375 or SK-MEL-28 cells treated with si-NC or ATG5 siRNA in combination with etoposide (n = 4). h Western blot of DDIT4 in A375 or SK-MEL-28 cells treated with si-NC or ATG5 siRNAs in combination with etoposide. DDIT4 band intensities normalized to si-NC + DMSO are displayed below the blots. i The illustration for the potential mechanism of autophagy inducer sensitizing drug response. b, d, f, g Data were presented as means ± SD. b, d, f The difference in multiple groups was estimated by one-way ANOVA analysis. g Two-sided student’s t test was used for the estimation of the difference in two groups. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001; ns is not significant. c, e, h See Supplementary Fig. 8 for uncropped data.