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. 2013 Oct 4;288(46):33171–33180. doi: 10.1074/jbc.M113.518175

FIGURE 7.

FIGURE 7.

Survivor TEL patch mutant cells have reduced expression of TPP1-FLAG, shTPP1, and GFP at later time points. A, Western blot analysis showing protein levels of TPP1-FLAG (anti-FLAG antibody) and loading control (β-actin) as a function of time in culture for the experiment shown in Fig. 1C. Samples from day 85 are derived from the experiment shown in Fig. 2A. B, quantitative RT-PCR analysis showing endogenous TPP1 mRNA levels in the indicated cell lines at the indicated time points. Endogenous TPP1 mRNA levels completely recovered after 96 days of culture. 18 S rRNA was used as the internal control during quantification. Error bars represent mean ± S.D.; n = 3. Reproducibility was high, so the error bars are difficult to see. Samples of days 7 and 38 are from the experiment shown in Fig. 1C, and samples of days 70 and 96 are from the experiment shown in Fig. 2A. C, flow cytometry analysis of GFP expressed from the stably integrated gene cassette (see Fig. 1A for a description of the gene cassette) in the indicated cell lines. GFP expression is no longer detectible in TEL mut1 and TEL mut2 cells that have been cultured for 60 days. All samples are derived from the experiment shown in Fig. 2A.

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