Extended Data Fig. 4: Manipulation of SAMHD1 levels by CRISPR/Cas9, shRNA, and lentiviral expression.
From: Thymidine nucleotide metabolism controls human telomere length
a, Immunoblot of 293T and K562 cells electroporated with Cas9 and the indicated sgRNA(s) using primary antibodies against SAMHD1 or β-Actin, corresponding to cell lines evaluated in Fig. 4b. b, Immunoblot of K562 cells transduced with vectors expressing the indicated shRNA, corresponding to cell lines evaluated in Fig. 4c. c, qRT-PCR of SAMHD1 expression compared to β-Actin, performed in technical triplicate. Means of the replicates are shown. d, Immunoblot of 293T cells transduced with vectors expressing the indicated shRNA. e, TRF of indicated cell lines transduced with the indicated shRNA and cultured for 15 days, and quantification of the TRF using the WALTER webtool. The boxplot displays the 75th, 50th and 25th percentile molecular weight of the telomere signal distribution in the TRF blot. f, Immunoblot of 293T cells transduced with vectors to overexpress either eGFP or the indicated SAMHD1 variant and treated with the indicated dose of dT, corresponding to cell lines evaluated in Fig. 4i, j. Data presented in this figure are the results of single experiments unless otherwise stated. Full-length western blots are presented as source data.
