Fig. 5: Experimental validation of TL regulation by perturbing expression of MMUT, RRM1, KIAA1429, and YWHAZ.
From: Unraveling the causal genes and transcriptomic determinants of human telomere length
A 34 days after HTR8/SVneo cells were infected with shRRM1 lentiviral particles, 8 shRRM1 monoclonal cell lines grown in 96-well plates were transferred to 6-well plates. TRF assay was used to measure the TL of these cell lines. B RRM1 RNA levels were estimated by qPCR and analyzed by GraphPad Prism software version 6.0. Data are represented as mean ± SD (n = 3). The two-sided P values of t test were (1.36e-7, 7.23e-8, 6.28e-7, 1.01e-5, 1.35e-7, 1.48e-7, 3.20e-7, 4.08e-7). ****P value < 0.0001. (C) HTR8/SVneo cells stably expressing control (shScramble) and shRRM1 were passaged over time (DAY) and examined for average TL by TRF (n = 4). D 34 days after HTR8/SVneo cells were infected with shMMUT lentiviral particles, six shMMUT monoclonal cell lines grown in 96-well plates were transferred to 6-well plates. TL of these six cell lines was measured by TRF assay. E MMUT RNA levels were estimated by qPCR and analyzed by GraphPad Prism software version 6.0. Data are represented as mean ± SD (n = 3). The two-sided P-values of t test were (1.23e-04, 2.34e-05, 2.05e-04, 2.19e-04, 7.63e-05, 1.78e-04). ***P value < 0.001; ****P value < 0.0001. F HTR8/SVneo cells stably expressing control (shScramble) and shRNA sequences against MMUT were passaged over time (DAY) and examined for average TL by TRF (n = 4). G 32 days after infection of shKIAA1429 lentiviral particles, six shKIAA1429 monoclonal cell lines from 96-well plates were grown in 6-well plates. The TL of these cell lines was measured by TRF assay. (H) KIAA1429 RNA levels were tested by qPCR and analyzed by GraphPad Prism software version 6.0. Data are represented as mean ± SD (n = 3). The two-sided P values of t test were (6.15e-05, 2.51e-05, 2.82e-05, 2.59e-05, 3.14e-05, 2.96e-05). ****P value < 0.0001. I 32 days after infection of shYWHAZ lentiviral particles, 5 shYWHAZ monoclonal cell lines grew from 96-well plates to 6-well plates. TL of these cell lines was measured by TRF assay. J YWHAZ RNA levels were estimated by qPCR and analyzed by GraphPad Prism software version 6.0. Data represent mean ± SD (n = 3). The two-sided P-values of t test were (7.80e-05, 7.80e-05, 7.30e-05, 7.70e-05, 1.01e-03). ****P-value < 0.0001.
