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. 2017 Jan 13;45(5):2615–2628. doi: 10.1093/nar/gkw1295

Figure 1.

Figure 1.

Telomerase inhibition in telomerase-positive cells onsets the alternative lengthening of telomeres to overcome the crisis. (A) Overview of ALT cell line generation from telomerase positive transformed cells (SW39). SW39 cells were introduced with Cas9/TERC targeting guide RNA and single cell clones were isolated and analyzed. (B) Terminal Restriction Fragment (TRF) analysis in SW39 (parental cell) and SW39 TERC KO survival clones (post-crisis cells). Digested DNA was run on 0.85% agarose gels. M indicates for the marker. (C) Droplet digital PCR based telomere repeat amplification (ddTRAP) analysis in SW39, SW39 TERC KO survival clones (SW39 ALT1, ALT2, and ALT3), SW26 and SW13 cells (mean ± SD; n = 3). (D) C-circle analysis in SW39 and SW39 ALT survival clones. One hundred nanograms of digested DNA from SW39 or SW39 ALT survival clones were used in the phi29 polymerase reaction.