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Additional file 1: Figure S1. of Mediator subunit Med12 contributes to the maintenance of neural stem cell identity

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posted on 2016-05-17, 05:00 authored by Nam Kim, Carolina Livi, P. Yew, Thomas Boyer
Unbiased genome-wide expression profiling analysis validation (A, C, D) RNA from mNS-5 NSCs infected with lentiviruses expressing non-specific (NS) control or Med12-specific shRNAs was harvested and subjected to RT-qPCR analysis. mRNA levels for each gene were normalized to β-actin mRNA and expressed relative to their corresponding mRNA levels in NS control shRNA-expressing cells. In (C, D) relative mRNA levels for 29 randomly selected genes from the original microarray-derived list were determined using the same RNA samples subjected to microarray analysis (C) as well as RNA from an independent set of knockdown experiments (D). Data represent the mean +/− SEM of at least three independent experiments performed in triplicate. Asterisks denote statistically significant differences relative to NS control shRNA (Student’s t-test, *p < 0.1, **p < 0.05, ***p < 0.01). (B) Nuclear extracts from mNS-5 NSCs infected with lentiviruses expressing non-specific (NS) control or Med12-specific shRNAs were harvested and resolved by SDS-10 % PAGE prior to processing by immunoblot using antibodies specific for Med12, Cdk8, the p89 subunit of transcription factor IIH (p89), and the β subunit of transcription factor IIE (TfIIeβ). Both p89 and TfIIeβ served as an internal loading controls. (TIF 16612 kb)

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National Institute of Mental Health

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