Fig. 2: TGS1 directs telomerase-dependent telomere maintenance involving Cajal bodies. | Nature Communications

Fig. 2: TGS1 directs telomerase-dependent telomere maintenance involving Cajal bodies.

From: TGS1 mediates 2,2,7-trimethyl guanosine capping of the human telomerase RNA to direct telomerase dependent telomere maintenance

Fig. 2

a Representative images of combined immunofluorescence with anti-TRF2 and anti-Coilin antibodies on H1299 cells transfected with indicated siRNAs for 10 days. b Percentage of Cajal bodies colocalizing with telomeres (TRF2) of the experiment shown in a. c Representative images of combined immunofluorescence with anti-TRF2 and anti-Coilin antibodies on H1299 cells treated, as described. d Percentage of Cajal bodies colocalizing with telomeres (TRF2) of the experiment shown in c. e–g Representative images of combined immunofluorescence with anti-TRF2 and anti-Coilin antibodies in tumor lung organoids treated as described. h Percentage of Cajal bodies colocalizing with telomeres (TRF2) on lung tumor organoids shown in e–g. i Representative images of combined immunofluorescence with anti-TRF1 and anti-FLAG antibodies from clone-22 cells transfected with indicated siRNAs. j Quantification of TRF1/FLAG-hTERT colocalization events of the experiment shown in i. k Representative images of hTR RNA-FISH combined with immunofluorescence using anti-TRF2 antibody on H1299 cells transfected with indicated siRNAs for 10 days. l Percentage of hTR colocalizing with telomeres (TRF2) per nucleus of cells shown in k. m Chromatin immunoprecipitation experiments (ChIP) using clone-22 cells treated, as described for 10 days. Mouse anti-TRF2 and mouse anti-FLAG antibodies were used. Mouse control IgM was used as negative control. Serial dilutions of chromatin extract (input) prepared from clone-22 cells, treated as described, were loaded. n Quantification of three independent ChIP experiments, average enrichment of telomeric repeats is indicated. For quantifications in b, d, h, j, l, and n, mean values are indicated; whiskers indicate standard deviation; N = number of independent experiments. n = number of analyzed nuclei. Arrowheads indicate colocalization events. Bars in panels b, d, j, l, and n indicate mean values. Red and green, epitopes detected by immunofluorescence; blue, DAPI-stained nuclei. Scale bars in a, c, i, k correspond to 1 µm. Scale bars in e–g correspond to 10 µm. A two-tailed Student’s t-test was used to calculate statistical significance; p-values are shown. Uncropped blots in Source Data.

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