Fig. 2: Mutations associated with Cartilage Hair Hypoplasia impair rRNA processing.
From: A disease-linked lncRNA mutation in RNase MRP inhibits ribosome synthesis
a Northern blot using ITS1 probe (probe 119) to detect pre-rRNA species in RNA extracted from CHH or healthy control (WT) fibroblasts. Pre-rRNA species indicated in green are present in both Pathway 1 and Pathway 2. 41 S in brown is a Pathway 1 intermediate, and those in blue are Pathway 2 species. b Quantification of relative abundance of 41 S to 47 S, and 30 S to 41 S, from northern blots as illustrated in a. Each dot represents an independent sample (total of 6 WT and 4 CHH samples; mean + /− SD). Includes samples processed in two independent northern blotting experiments. Indicated p value for 30/41 S derived from two-tailed t test (t = 3.463, df=8). c Northern blot using probes against ITS1 (left; probe 119) and ITS2 (right; probe 123), to detect pre-rRNA in parental K562 cells, or CRISPR-generated clones of K562 with a 70AG mutation in RMRP. C, D, and F represent cell lines derived from independent CRISPR clones. d Quantification of pre-rRNA species from WT and 70AG K562 cells. The intensity of bands for each indicated species were first normalized to the 47 S band in that lane. Values from WT cells were then subtracted from mutant values. Includes data from three independent experiments (mean + /− SD). e Northern blot with probe against 5.8 S rRNA, showing the short (5.8SS) and long (5.8SL) form of these species in indicated cell lines. f Quantification of ratio between 5.8SS and 5.8SL, determined from northern blots illustrated in e. Includes data from three independent rounds of RNA extraction, run on two separate gels, including total of 4 wild type and 8 mutant samples (mean + /− SD). Source data are provided as a Source Data file.
