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. 2021 Mar 29;12(4):334. doi: 10.1038/s41419-021-03574-2

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© The Author(s) 2021

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. 2 — A qRT-PCR assay was performed to detect the silencing effect of ATF4 siRNA in PANC-1 and BxPC-3 cells. n = 3, ^***p < 0.001. B, C CCK8 analysis showed the growth curve of PANC-1 and BxPC-3 cells transfected with ATF siRNA. n = 3, ^**p < 0.01, ^***p < 0.001. D, E Plate colony formation assays showed that colony formation decreased in PANC-1 and BxPC-3 cells with ATF4 silencing. n = 3, ^**p < 0.01, ^***p < 0.001. F, G. Transwell assays showed the fractions of migrated and invaded PANC-1 and BxPC-3 cells. Silencing ATF4 expression decreased the migration of invaded pancreatic cancer cells. n = 3, ^**p < 0.01, ^***p < 0.001. H Image of harvested tumors from PANC-1 cells with stable ATF4 knockdown or mock cells subcutaneously transplanted into the bilateral hind legs of nude mice. I The tumor weights are shown as the mean ± SD, n = 5. ^*p < 0.05. J The tumor growth curve. The points and bars represent the mean ± SD. n = 5, ns not significantly different. ^*p < 0.05. K qRT-PCR assay analyzed the mRNA expression level of ATF4 in tumor tissues from shATF4 PANC-1 cells compared with mock cells. n = 5, ^***p < 0.001. L, M IHC staining showed that the positive expression rate of Ki67 was lower in tumor samples from shATF4 PANC-1 cells than in mock cells. n = 5, ^***p < 0.001.