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. 2024 Sep 14;15:8044. doi: 10.1038/s41467-024-51747-2

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© The Author(s) 2024

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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Fig. 2 — a Flow cytometry analysis of HaCaT cells stained with anti-DSG3 antibodies or IgG as control. HaCaT cells were transduced with sgNT, sgKLF5_1 (p = 0.0698), or sgKLF5_2 (p = 0.0206). A representative histogram and mean fluorescence intensity of four independent experiments are displayed. b Western blot analysis of HaCaT cell lysates using KLF5, DSG3, and GAPDH antibodies. HaCaT cells were stably transduced with sgNT, sgKLF5_1, or sgKLF5_2. Representative Western blot images and quantifications of the respective proteins are shown; sgKLF_1 (p = 0.0005, n = 7), sgKLF5_2 (p = 0.0023, n = 6). Values were normalized to sgNT. c Quantitative Real-time PCR analysis of DSG3 of mRNA extracted from HaCaT cells stably transduced with sgNT, sgKLF5_1, or sgKLF5_2. Values were normalized to sgNT. n = 6, sgKLF5_1 p = 0.0235; sgKLF5_2 p < 0.0001. d Immunofluorescence staining of HaCaT cells stably transduced with sgNT, sgKLF5_1 or sgKLF5_2 using DSG3 antibodies and DAPI as nuclear stain (n = 3, sgKLF5_1 p < 0.0001, sgKLF5_2 p = 0.0009). Representative pictures are shown. Scale bar = 10 μm. e Dispase-based dissociation assay of HaCaT cells stably transduced with sgNT, sgKLF5_1, or sgKLF5_2 (n = 4, sgKLF5_1 p = 0.0046; sgKLF5_2 p = 0.0020). Representative images and the number of fragments are shown. f Western blot analysis of HaCaT cell lysates stably overexpressing KLF5 using KLF5, DSG3, and GAPDH antibodies. Representative Western blot images and quantifications of respective proteins (n = 7, KLF5 p = 0.0006, DSG3 p = 0.0555), are shown. g Dispase-based dissociation assay of HaCaT cells stably overexpressing KLF5 (n = 4, p = 0.0031). Representative images and the number of fragments are shown. Values expressed as mean with standard deviation (mean ± SD). One n represents one biological replicate. Source data are provided as a Source Data file. Experiments (a–c, f) were analyzed with one-sample t-test (two-sided), (d, e) were analyzed with One-way-ANOVA, SIDAK correction. g was analyzed with student’s t-test (two-sided). p < 0.05*; p < 0.01**; p < 0.001***.