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. 2024 Mar 16;15:2384. doi: 10.1038/s41467-024-46602-3

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© The Author(s) 2024, corrected publication 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. 7 — a Immunoblotting of Cas9 and MCP in RAW264.7 cells transduced with lenti-dCas9-VP64 and lenti-MS2-P65-HSF1. b qPCR (upper) and immunoblotting (lower) of Tead3 in RAW264.7 cells with CRISPRa-mediated overexpression of Tead3. n = 3 biological replicates per group. c, d TRAP staining images (c) and quantification (d) of control and Tead3-overexpressing RAW264.7 cells treated with RANKL (50 ng/mL) for 5 days. Multinucleated TRAP-positive cells (outlined by dashed lines) were counted. Scale bars, 100 μm. n = 3 wells per group. e–g qPCR of Nfatc1 (e), Trap5 (f), and Ctsk (g) in control and Tead3-overexpressing RAW264.7 cells treated with RANKL (50 ng/mL) for 3 days. n = 3 biological replicates per group. h Immunoblotting of Nfatc1, Ctsk, and Hsp90 in control and Tead3-overexpressing RAW264.7 cells treated with RANKL (50 ng/mL) for 2 days and 5 days. i Immunoblotting of Tead3 and Hsp90 in RAW264.7 cells transfected with two independent Tead siRNAs or scrambled negative control (NC). j, k TRAP staining images (j) and quantification (k) of control and Tead3-knockdown RAW264.7 cells treated with RANKL (50 ng/mL) for 5 days. Multinucleated TRAP-positive cells (outlined by dashed lines) were counted. Scale bars: 100 μm. n = 3 wells per group. l Immunoblotting of Nfatc1, Ctsk, and β-actin in control and Tead3-knockdown RAW264.7 cells treated with RANKL for 3 days and 5 days. m Immunoblotting of Tead3 and Hsp90 in control and Malat1-knockdown RAW264.7 cells transfected with Tead3 siRNA or scrambled negative control (NC). n–p Control and Malat1-knockdown RAW264.7 cells were transfected with Tead3 siRNA or scrambled negative control (NC). 24 h after siRNA transfection, the cells were treated with RANKL for 5 days, followed by TRAP staining (n) and quantification (o) of multinucleated TRAP-positive cells (outlined by dashed lines). Scale bars, 100 μm. Cell lysates were subjected to immunoblotting of Nfatc1, Ctsk, and β-actin (p). n = 3 wells per group in o. Statistical significance in b, d–g, k, and o was determined by a two-tailed unpaired t test. Error bars are s.e.m. The experiments in a, h, i, l, m, and p were repeated independently three times, yielding similar results. Source data are provided as a Source Data file.