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. Author manuscript; available in PMC: 2016 Mar 24.
Published in final edited form as: Nature. 2015 Sep 24;525(7570):523–527. doi: 10.1038/nature14978

Extended Data Figure 10. KBTBD8 controls translation.

Extended Data Figure 10

a. hESCs stably depleted of KBTBD8 were subjected to neural conversion for 3 days, and hESCs and differentiating cells were analyzed by RNA deep sequencing and ribosomal profiling to determine translation efficiency. Distribution of translation efficiency changes for 7725 mRNAs brought about by KBTBD8 depletion is shown. b. hESCs stably depleted of either TCOF1 or KBTBD8 were subjected to neural conversion for 3 days, and translation efficiency was determined by RNAseq and ribosome profiling. c. Translation efficiency blot of differentiating hESCs transduced with control- or KBTBD8-shRNAs were labeled for significantly affected transcripts in general (blue), with links to CNS precursor formation (gold), or with links to neural crest formation (green). d. hESCs stably depleted of KBTBD8 or TCOF1 were subjected to neural conversion for 3 days, and expression levels of indicated proteins were analyzed by Western. e. hESCs stably depleted of KBTBD8 were subjected to neural conversion for 3 days, and levels of ATRX1 and PCM1 mRNA were determined by qRT-PCR (mean of 3 technical replicates +/− s.e.m.). f. hESCs stably depleted of KBTBD8 were subjected to neural conversion for 3 days, and protein stability of ATRX1 and PCM1 was determined by cycloheximide chase and Western (mean of 3 biological replicates +/− s.d., ATRX1 and PCM1 levels were normalized relative to actin levels and 0h time point set to 100%).