Extended Data Figure 6. Footprinting validation of a RiboSNitch in 5’UTR of MRPS21 identified by PARS.
a, Gel analysis of 150mer fragments of MRPS21 RNA using S1 nuclease (lanes 5 (Father), 6 (Mother)), and SHAPE probing ((lanes 9 (Father), 10 (Mother)). Additionally, sequencing lanes (lanes 1,2), uncut (lane 3 (Father), lane 4 (Mother), and DMSO treated lanes (lane 7 (Father), lane 8, (Mother)) are also shown. Black arrows indicate the change in structure between the Father and Mother alleles. b, Top: The sequence of a portion of the transcript containing the RiboSNitch was shown. The RiboSNitch is in red. Bottom: Single strand profile by S1 sequencing of the father and mother allele. Y axis indicates the percentage of signal at each base over the total signal in the region. c,d, SAFA quantification of manual structure probing of both MRPS21 alleles using S1 nuclease (c) and SHAPE (d). e, S1 sequencing reads are mapped uniquely to either the A or C allele in the child. The grey box indicates the bases that show structural differences by allele speficic mapping in the child. f, Gel analysis of 150mer fragments of MRPS21 RNA using DMS footprinting (lanes 1,2 and 3 (Father), 4, 5 and 6 (Mother)). Black arrows indicate the change in structure between Father and Mother alleles. g, Quantification of DMS footprinting of both MRPS21 alleles using SAFA.