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. 2018 Nov 14;12(12):2165–2181. doi: 10.1002/1878-0261.12392

Figure 1.

Figure 1

The HDAC4 expression in human fibroblasts. (A) Schematic representation of the different HDAC4 versions used in this study. Phosphorylated serine residues, binding sites for 14‐3‐3 proteins are indicated. (B) Immunoblot analysis using the indicated antibodies in BJ/TERT expressing the indicated transgenes. Vimentin was used as loading control. (C) Normal human diploid fibroblasts (BJ) expressing the telomerase catalytic subunit (TERT) infected with retrovirus expressing the indicated genes and stained for senescence‐associated β‐galactosidase (SA‐β‐gal) marker. Scale bar: 50 μm. (D) Quantitative analysis of SA‐β‐gal positivity from experiments described in (C). Data are expressed as means ± SD, n = 3. Student t‐test: *P < 0.05, ***P < 0.005. (E) Immunoblot analysis using the indicated antibodies in BJ/TERT expressing the indicated transgenes following treatment with 4‐OHT for 48 h. RACK1 was used as loading control. (F) Analysis of cells synthesizing DNA, as scored after BrdU staining at the indicated times. Data are expressed as means ± SD, n = 4. Student t‐test: **P < 0.01, ***P < 0.005. (G) BJ‐TERT expressing the indicated transgenes after 8 days of induction were stained for SA‐β‐gal activity. Scale bar: 50 μm. (H) Quantitative analysis of SA‐β‐gal positivity from experiments described in (G). Data are expressed as means ± SD, n = 4. ***P < 0.005. (I) Immunoblot analysis using the indicated antibodies in BJ/TERT expressing the indicated transgenes following treatment with doxycycline for 48 h. Vimentin was used as loading control. (J) Quantitative analysis of SA‐β‐gal positivity in BJ‐TERT cells expressing doxycycline‐inducible versions of the indicated transgenes after 8 days of induction. Data are expressed as means ± SD, n = 3. Student t‐test: *P < 0.05, **P < 0.01, ***P < 0.005.