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. 2016 Apr 1;36(8):1297–1309. doi: 10.1128/MCB.00950-15

FIG 5.

FIG 5

Cellular stretching induces nuclear translocation of Gtf2ird1. (A) Immunocytochemistry of a control rat tenocyte without stretching. Gtf2ird1 was seen predominantly within the cytoplasm in comparison with localization in the nucleus (green, Gtf2ird1; blue, Hoechst). (B) Immunocytochemistry of a rat tenocyte after cellular stretching. Gtf2ird1 expression was most concentrated in the nucleus and also moderately concentrated in the cytoplasm surrounding the nucleus (green, Gtf2ird1; blue, Hoechst). (C) Proportion of cells with nuclear translocation induced by stretching. Error bars represent standard errors of the means (***, P < 0.001, two-tailed Student's t test). (D) Western blot of fractionated protein extracted from stretched tenocytes confirms that Gtf2ird1 translocates into the nucleus. α/β-Tubulin and histone H3 were used as cytoplasmic and nuclear controls, respectively. (E) Gtf2ird1 knockdown using siRNA (siGtf2ird1) inhibits Mkx expression despite stretching, indicating the importance of Gtf2ird1 in mechanosensitive Mkx regulation. Error bars represent standard errors of the means (*, P < 0.05, ***P < 0.001, two-tailed Student's t test).