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. 2013 Mar 4;200(5):605–617. doi: 10.1083/jcb.201206121

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© 2013 Dreesen et al.

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Figure 1. — Loss of LMNB1 in senescent human keratinocytes. (a and b) Immunofluorescence staining of primary keratinocytes (N 50; a) and hTERT-positive keratinocytes (N/TERT-1; b) with LMNA/C (LA/C), LMNB1 (LB1), and LAP2 antibodies. Merged images are shown for N/TERT-1 cells only. (right) Phase contrast (PH) images are shown for N 50. Arrowheads show large flattened cells. Bars, 10 µm. (c) SA-β-gal staining of hTERT-positive (N/TERT-1) and primary (N 50) keratinocytes at PD 28 and 21, respectively. Same fields shown in phase contrast (top) and bright field (bottom). The percentages of SA-β-gal–positive cells are indicated. Bars, 20 µm. (d) LMNB1, LMNA/C, and LAP2-α levels during a 28 PD time course of primary N 50 cells (left) and one independently passaged N/TERT-1 line (right) by Western blotting. Quantified signal intensities of LMNB1, LMNA/C, and LAP2-α and normalized to GAPDH control are plotted below (n = 3). The results are presented as the means ± SD of three independent experiments.