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. 2021 Mar 12;4:332. doi: 10.1038/s42003-021-01857-0

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© The Author(s) 2021

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 — a TGF-β2 induced expression of Prg4 in superficial zone articular chondrocytes is blocked by inhibition of p38 activity. SZCs were cultured in either the absence (white) or presence (gray) of TGF-β2 (20 ng/ml), plus a p38 inhibitor (SB203580; 10 μM), or a JNK inhibitor (SP600125; 10 μM), as indicated for 2 days. Gene expression was assayed by RT-qPCR and normalized to Gapdh. Error bar indicates standard error of the mean (n = 4 technical repeats for Prg4 expression, and n = 2 technical repeats for Creb5 expression). Similar results have been obtained in three independent biological repeats. b, c DZCs were infected with a lentivirus encoding doxycycline-inducible Creb5-HA. After selection in G418, the cells (iCreb5-DZCs) were cultured in either the absence or presence of doxycycline (1 μg/ml), TGF-β2 (20 ng/ml), a p38 inhibitor (SB203580; 10 μM), or a JNK inhibitor (SP600125; 10 μM), as indicated for 2 days. b Gene expression was assayed by RT-qPCR and normalized to Gapdh. Error bar indicates standard error of the mean (n = 2 technical repeats). Similar results have been obtained in three independent biological repeats. c Western analysis of exogenous Creb5-HA expression and phosphorylation in iCreb5-DZCs with antibodies directed against either phospho-Creb5 (T61), total Creb5, HA, or α-tubulin. Similar results have been obtained in two independent biological repeats. d, e DZCs were infected with a lentivirus encoding either iCreb5-WT-HA; iCreb5-T59,61A-HA; or iCreb5-T59,61D-HA. After selection in G418, the cells were cultured in either the absence or presence of doxycycline (1 μg/ml), TGF-β2 (20 ng/ml), and TGF-α (100 ng/ml) as indicated for 3 days. d Western analysis of iCreb5-WT/mutant expression in iCreb5-DZCs with antibodies directed against either phospho-Creb5 (T61) or total Creb5. Similar results have been obtained in two independent biological repeats. e Gene expression was assayed by RT-qPCR and normalized to Gapdh. Error bar indicates standard error of the mean (n = 2 technical repeats). Similar results have been obtained in three independent biological repeats.