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. 2015 Jun 8;11(6):e1005286. doi: 10.1371/journal.pgen.1005286

Fig 5. Endogenous modulation of hTERT intron 4 alternative splicing affects telomerase activity.

Fig 5

(A) Schematic of position of binding of a morpholino (Ex4/Int4) (purple line) utilized to block splicing at the exon 4/intron 4 junction. (B) RT-PCR to determine concentration-dependent changes in INS1b and FL-hTERT transcript levels after delivery of the Ex4/Int4 morpholino into HEK293T cells. (C) Direct telomerase activity assay on immunopurified telomerase from equal numbers of HEK293T cells treated with 10 μM standard control (Ctrl) or Ex4/Int4 morpholino for 48 hours. (D) Quantification of the direct telomerase activity assay normalized to standard control morpholino (mean ± SEM; n = 3; P-value calculated by two-tailed Student’s t test; *p≤0.05). (E) Quantification of INS1b and FL-TERT expression after 10 μM morpholino treatment determined by RT-PCR on cells from each biological replicate of morpholino treatment. The INS1b:hTERT transcript levels ratio was calculated by densitometry and normalized to standard control morpholino (mean ± SEM; n = 3; P-value calculated by two-tailed Student’s t test; *p≤0.05).