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. 2023 Mar 3;14:1214. doi: 10.1038/s41467-023-36881-7

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© The Author(s) 2023

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. 3 — Enhanced Cd276 promoter activity in Tsc2−/− 105K cells expressing empty vector (EV) compared to reconstitution of TSC2 (a) and in Tsc2 KO MEFs compared to Tsc2-WT MEFs (b). Relative luciferase activity was determined by a dual-luciferase assay system. psiCHECK2-Cd276 encodes the Cd276 promoter. Empty psiCHECK2 vector was used as the negative control. n = 6, means ± SD, two-way ANOVA with Holm-Sidak’s multiple comparisons test, *p < 0.05, ***p < 0.001. Raptor, mTOR or S6K knockdown suppresses Cd276 promoter activity in Tsc2−/− 105K cells (c) and Tsc2 KO MEFs (d). n = 6, means ± SD, two-way ANOVA with Holm-Sidak’s multiple comparisons test, *p < 0.05, **p < 0.01, ****p < 0.0001. siRNA knockdown of YY2 reduces B7-H3 protein expression in Tsc2−/− 105K cells (e) and Tsc2 KO MEFs (f) (n = 3). Knockdown of YY2 reduces Cd276 mRNA expression in Tsc2−/− 105K cells (g) and Tsc2 KO MEFs (h). n = 3, means ± SD, two-tailed unpaired Student’s t-test, *p < 0.05, ***p < 0.001. i Promoter region of Cd276 displaying the location of the YY2 binding site. YY2 ChIP-qPCR analysis showing increased YY2 occupancy on the Cd276 promoter in Tsc2−/− 105K cells (j) and Tsc2 KO MEFs (k). n = 4, means ± SEM, two-way ANOVA with Holm-Sidak’s multiple comparisons test, *p < 0.05, **p < 0.01, ****p < 0.0001. Cd276 promoter activity is suppressed by YY2 knockdown in Tsc2−/− 105K cells (l) and Tsc2 KO MEFs (m). n = 6, means ± SD, two-way ANOVA with Holm-Sidak’s multiple comparisons test, *p < 0.05, ***p < 0.001. Source data and exact p values are provided in the Source data file.