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. 2019 Apr;25(4):496–506. doi: 10.1261/rna.067967.118

FIGURE 1.

FIGURE 1.

Effects of Bcd1p depletion on RNA expression level and RNA polymerase II recruitment at selected gene loci. (A) Yeast GAL::3HA-BCD1 cells were shifted from galactose YPG medium to glucose YPD medium for the times indicated before total protein extraction and western blotting. The Bcd1p signal was monitored using an anti-HA antibody. The aspartyl-tRNA synthetase Dps1p was used to control protein loading. (B) Scheme of the RT-qPCR strategy that allowed quantification of total (mature plus immature) or immature snoRNA expression levels. (C,D) Cultures of yeast GAL::3HA-BCD1 cells were maintained in YPG medium or shifted to YPD medium for 6 h (C) or 16 h (D) before RT-qPCR assays. Relative RNA expression level in YPG condition was set to one. The ALG9 gene coding mannosyltransferase was used as an endogenous control. (E) Cultures of yeast RPB1-TAP x GAL::3HA-BCD1 cells were maintained in YPG medium or shifted to YPD medium for 16 h before chromatin immunoprecipitation (ChIP) assays. IP was performed using IgG-Sepharose beads and the GAL::3HA-BCD1 strain was used as a control.