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. 2009 Jun 1;185(5):797–810. doi: 10.1083/jcb.200810084

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© 2009 Jiang et al.

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.jcb.org/misc/terms.shtml). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/).

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Figure 4. — Induction of APBs in SV40-immortalized ALT cells by siRNA-mediated knockdown of LTAg. (A) Western blots showed decreased LTAg in both IIICF-T/B3 (containing wt LTAg and one wt TP53 allele) and IIICF-402DE/D2 (containing mutant LTAg that does not bind p53, and no wt TP53 alleles) cells 48 h after SV40T siRNA transfection. Treatment with SV40T siRNA induced p21 in IIICF-T/B3 but not in IIICF-402DE/D2 cells. The blots were probed with the indicated antibodies. (B) Double immunofluorescence of SV40T and p21 in IIICF-T/B3 and IIICF-402DE/D2 cells treated with SV40T or control siRNAs for 4 d. Strong p21 staining was detected in IIICF-T/B3 cells depleted of SV40T. (C) IIICF-T/B3 cells were triple stained for TRF1, p21, and SV40T 4 d after SV40T siRNA transfection. APBs (visualized here as large TRF1 foci) were observed in cells with high levels of p21. Bars: (B) 100 µm; (C) 20 µm.