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. 2019 Jul 22;10:3279. doi: 10.1038/s41467-019-11111-1

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© The Author(s) 2019

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. 5 — PP1α-Cavin3 complex in UV-induced DNA damage response. a PLA interaction between Cavin3 and PP1α upon UV treatment. DAPI detects the nucleus (blue). b Quantification of PLA Cavin3/PP1α interactions in control and UV-treated cells as the mean ± SD, three individual experiments. ****p-value (Student’s t-test, two tail) < 0.0001. c The number and ratio of nuclear (Nuc.; purple) and cytoplasmic (Cyto.; gray) localized PLA signals for Cavin3-PP1α interactions presented as mean ± SD, three independent experiments in a histogram and table. d Quantitation of caveolae number in untreated and UV-treated A431 cells. Caveolae number on the apical surface from four sets of images were quantified blinded from three independent experiments and were normalized to the length of the surface sampled (over 150 μm for each set of images where **p = 0.0027, Student’s t-test, two tail). e A431 cells transfected with scrambled siRNA (control), Cavin3 and PP1α individually or together were exposed to UV treatment. Lysates were western blotted for γH2AX and H2AX anti-PP1α and anti-Cavin3 and α-Tubulin antibodies. f Relative protein expression (%) of γH2AX (Scrambled vs siCavin3: *p = 0.0172, Scrambled vs siPP1: ****p < 0.0001), Cavin3 (Scrambled vs siCavin3: ****p < 0.0001, Scrambled vs siCavin3+siPP1: ****p < 0.0001), and PP1α (Scrambled vs siPP1: ****p < 0.0001, Scrambled vs siCavin3+siPP1: ****p < 0.0001), three independent experiments, one-way ANOVA with a Tukey’s multiple comparison test. g Western blot analysis of cleaved PARP, cleaved-caspase-3, total caspase-3 and GAPDH in PP1α- or/and Cavin3-knockdown A431 cells. h Relative protein expression (%) of cleaved PARP (Scrambled vs siCavin3: *p = 0.0486, Scrambled vs siPP1: ****p<0.0001), cleaved caspase-3 (Scrambled vs siCavin3: *p = 0.0465, Scrambled vs siPP1: ***p = 0.0001) and total caspase-3 (Scrambled vs siCavin3: *p = 0.0109, Scrambled vs siPP1: ****p < 0.0001). Three independent experiments analyzed using one-way ANOVA. i LDH release (% of total LDH) was calculated (Scrambled vs siPP1: ****p < 0.0001, Scrambled vs siCavin3: *p = 0.0243). Significance was determined by one-way ANOVA and a Bonferroni test from three independent experiments