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. 2021 Jan 29;12:674. doi: 10.1038/s41467-020-20632-z

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© The Author(s) 2021, corrected publication 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. 1 — a Adenoviral Neurog3, Pdx1 and Mafa in islets (inset, endogenous gene expression) (n = 5 animals; paired t-test). b Islets transduced with Ad-M3C (β-cell mature; B-MAT) lose β-cells occupying the bottom 15 percentile for PDX1 compared to controls (β-cell normal; B-NORM) (inset, non-normalized polynomial-fitted B-NORM distribution) (n = 6 islets/3 animals; two-way ANOVA, Bonferonni’s multiple comparison) (F = 18.75, DF = 20). c As for b, but showing the frequency distribution for MAFA (n = 8 islets/3 animals; two-way ANOVA, Bonferonni’s multiple comparison) (F = 3.03, DF = 20). d Images showing more homogenous PDX1/MAFA fluorescence in B-MAT islets (scale bar = 60 µm). e–g INS-PDX1 (e), INS-MAFA (f) and MAFA-PDX1 (g) are positively correlated (n = 137 cells, linear regression). h The linear correlation between PDX1 and BFP in Pdx1-BFP islets is lost following Ad-M3C transduction (B-MAT) (n = 465 cells/3 animals). i BFP^LOW cells (prior immature) become PDX1^HIGH in B-MAT islets, while BFP^HIGH cells (prior mature) remain PDX1^HIGH (n = 93 cells/3 animals; one-way ANOVA with Sidak’s multiple comparison) (F = 52.12, DF = 3). j Images from Pdx1-BFP islets showing cells that underwent PDX1^LOW - > PDX1^HIGH conversion (arrow shows a cell that remained PDX1^HIGH) (scale bar = 50 µm) (n = 5 islets/3 animals; two-way ANOVA, Bonferroni’s multiple comparison) (F = 2.80, DF = 18). k–q No differences are detected in the ratios of α- to β-cells (n = 23 islets/3 animals) and δ- to β-cells (n = 18 islets/3 animals) (k–n), or the proportion of PDX1 + /INS− or PDX1 + /GLU + cells (n = 10 islets/4 animals) (o–q) in B-MAT islets (unpaired t-test) (scale bar = 40 µm). r No difference in TUNEL+ cell numbers is detected in B-MAT islets (n = 18 islets/4 animals; unpaired t-test) (scale bar = 42.5 µm). s Cell proliferation is similar in B-NORM and B-MAT islets, as shown by PCNA staining (n = 24 islets/4 animals; unpaired t-test) (scale bar = 42.5 µm). t Transition to high PDX1/MAFA content occurs in PDX1^LOW/MAFA^LOW cells (1), whereas PDX1^HIGH/MAFA^HIGH cells remain unaffected (2), with PDX1/MAFA levels never surpassing those in B-NORM islets (3). Bar graphs show the mean ± SEM. Violin plot shows median and interquartile range. Box-and-whiskers plot shows median and min-max. All tests are two-sided where relevant. BFP-blue fluorescent protein; INS-insulin; GLU-glucagon; SST-somatostatin; TUNEL-terminal deoxynucleotidyl transferase dUTP nick-end labeling; PCNA-proliferating cell nuclear antigen.