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. 2020 Feb 14;11:911. doi: 10.1038/s41467-020-14684-4

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© This is a U.S. government work and not under copyright protection in the U.S.; foreign copyright protection may apply 2020

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. 4 — a FLAG tagged or 3xTy1 tagged CASZ1b is stably cloned into RD and SMS-CTR cells. CASZ1b expression is Tetracycline (Tet) or Doxycycline (Dox) inducible shown by western blot. b Restoration of CASZ1b in RD cells (Tet treatment) or SMS-CTR cells (Dox treatment) inhibits cell proliferation compared to control cells (Ctrl) shown by IncuCyte cell confluence assay. c Restoration of CASZ1b in RD cells increases expression of skeletal muscle differentiation markers MYOG and MHC protein shown by western blot. d Staining of SMS-CTRtetCASZ1b cells with Phalloidin (red) and DAPI (blue) shows that cells with restoration of CASZ1b (Dox + ), but not control (Dox-) form multinuclear cells (middle panel, red arrow). Bottom panel shows the enlarged image from the middle panel, and the arrow with the same color indicates these nuclei are in one cell. e–h GSEA assays of RNA-seq data show the positive enrichment of MYOD signature genes and human skeletal muscle differentiation genes after the restoration of CASZ1b in SMS-CTR cells, and negative enrichment of Rb repressed genes and E2F target genes. Data represent mean ± SEM for technical triplicates, *p < 0.01. Two-sided Student’s t-test was used to calculate statistical difference. Source data are provided as a Source Data file.