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. 2022 Dec 13;12:21496. doi: 10.1038/s41598-022-25405-w

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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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Islet cell differentiation and characterization in PANC-1 cells using PJ34 mediated PARP1 inhibition. (a) Phase contrast microscopic images of PANC-1 cells treated with PARP1 inhibitor PJ34 at day 0, 3rd and 10th. Graph show quantification of cell aggregation or clusters generated per field of image from PANC-1 cells with PJ34 treatment at 10 days of treatment. (b) Immunoblot profiling for PARP1 protein following PJ34 inhibition at short and long-term exposure. (c) Phase contrast microscopic images of PANC-1 cells differentiated with control (SFM), activin-A, and activin-A with PJ34 in combination at day 10^th of differentiation process. Color brightfield images show presence of brick-red insulin staining after differentiation using dithizone (DTZ) stain. (d) Quantification for islet-like clusters generated per field of image from PANC-1 cells in control, activin-A and activin-A with PJ34 treatment after 10 days of differentiation. (e) Immunocytochemical characterization of generated clustered from PANC-1 cells followed activin-a and PJ34 treatment for insulin (green) to confirm β cell differentiation. Graph show quantification for mean fluorescence intensity for human insulin production in differentiated cells. Data represented as mean ± sem, p-value show significance at 95% confidence interval using two-way anova analysis compared to activin-A differentiated cells (n = 3). (f) Quantification and modelling for presence of insulin in activin-a differentiated cells with and without PJ34 using 2-dimensional mean fluorescent intensity dot plot representation extracted from immunostained images with Axio-vision Zen 10 software, Zeiss, Canada, where insulin stays on Y-axis and dapi is marked on x-axis.