Extended Data Fig. 1: ATI of DGCR8 links dynamic Microprocessor autoregulation to altered DGCR8:DROSHA stoichiometry.

a, Ratio of splicing events covering different exons based on RNA-seq data during mES cell-to-EB differentiation over a 13-day time course (Fig. 1a). b, Agarose gel analysis of 5′ RACE products (left) and summary of Sanger sequencing of 5′ RACE colonies (right). PCR products outlined by red arrows were purified and further processed for cloning and Sanger sequencing. The numbers indicate the proportion of all sequenced clones that map to a particular nucleotide. Stem-loop 1 (SL1) sequence in 5′ UTR is highlighted in red (Fig. 1b). c, Stem-loop structures (SL1 and SL2) in the 5′ UTR and CDS region of DGCR8 mRNA. Green arrowheads indicate CRISPR–Cas9 design for SL1 deletion. d, PCR analysis of genomic DNA for SL1 knockout (ΔSL1) in mES cells. e, qRT–PCR analysis of DGCR8 and DROSHA mRNA expression in wild-type and ΔSL1 mES cells. Data were normalized to GAPDH, and error bars indicate s.d. (n = 3, technical replicates). f, Western blots of DGCR8 and DROSHA proteins in wild-type and ΔSL1 mES cells transfected with corresponding siRNAs. Experiments were repeated three times with similar results (Methods).