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. 2021 Feb 24;40(11):2065–2080. doi: 10.1038/s41388-021-01664-1

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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 Total extracts of the non-transduced PANC-1 cell line, of the PANC-1 transduced with shRNA directed against GAPDH, and the KD clones PZT-2 and PZT-1 were analyzed by western blot using an anti-H2A.Z antibody (n = 3). Actin was used as loading control. b Expression of H2A.Z.1, H2A.Z.2.1, and H2A.Z.2.2 were analyzed by RT-qPCR in the KD clones PZT-2 and PZT-1. Values are normalized to the expression in the untransduced PANC-1 cell line. GAPDH was used as house-keeping gene. The means ± SEM of three biological replicates is shown. Significance was analyzed using two-way ANOVA: *p value ≤ 0.05; **p value ≤ 0.01; ***p value ≤ 0.001. c Total extracts of the PANC-1 cell line, the PGAPDH cell line and the clones PZT-2 and PZT-1 were analyzed by western blot to identify expression and cleavage of caspase 3, PARP and survivin (n = 3). d PANC-1, PGAPDH, PZT-2, and PZT-1 cell lines present low caspase 3 and 7 activity with respect to C-33A cells in which apoptosis was induced by UV light, n = 3. Significance was analyzed by one-way ANOVA. ****p value ≤ 0.0001. e Cell growth curves of PANC-1, PGAPDH, PZT-2, and PZT-1. The means ± SEM of three biological replicates is shown. Significance was analyzed by two-way ANOVA. ***p value = 0.001; ****p value ≤ 0.0001. f Clonogenic capacity of the cell lines PANC-1, PGAPDH, PZT-1, and PZT-2. The graph shows absorbance at 590 nm of the crystal violet retained in the colonies. The means ± SEM of three biological replicates is shown. Significance was analyzed by one-way ANOVA. ****p value ≤ 0.0001. g Incorporation of BrdU into the cell lines PANC-1, PGAPDH, PZT-2, and PZT-1 24 h after synchronization in G0 by serum starvation for 48 h. The means ± SEM of three biological replicates is shown. Significance was analyzed by one-way ANOVA. *p value = 0.0497. h Distribution of the cell line PANC-1, PGAPDH, PZT-2, and PZT-1 in cell cycle phases G0–G1, S and G2-M as examined by staining with BrdU and IP 24 h after synchronization in G0 by serum starvation for 48 h. The means ± SEM of three biological replicates is shown. Significance was analyzed by two-way ANOVA. *p value = 0.0415; ****p value ≤ 0.0001.