Fig. 1
From: Detecting RNA base methylations in single cells by in situ hybridization
Destabilized base pairing by methylated RNA enables detection of post-transcriptional modification in single cells. a N6-Dimethylation of A1518 and A1519 in 16S rRNA catalyzed by KsgA. b N6-Dimethyladenine (m62A) is sterically inhibited from base pairing with thymine. c A Molecular Beacon can discriminate between methylated and unmethylated synthetic RNA, as revealed by in vitro thermal melting. d MR-FISH assay for rRNA methylation, using two Molecular Beacons and two strains of E. coli:, one expressing active KsgA (parent strain) and one mutant strain with the methyltransferase deleted from its genome (ΔksgA). e Fluorescence images showing that ΔksgA E. coli are stained red and green by MR-FISH probes, while parent strain cells are brightly stained only by the methylation-insensitive red probe. Scale bar: 10 μm. f Scatter plots of red and green fluorescence intensities of parent strain (n = 1315 cells) and ΔksgA (n = 1112 cells) bacteria stained by MR-FISH. g Log-normally distributed ratios of red fluorescence intensity (IR) to green fluorescence intensity (IG) of bacteria stained by MR-FISH. h Box plots (box: 25th–75th percentiles; whiskers: 5th–95th percentiles; horizontal line: medians; squares: means) of two-color ratios shown in (g) (empty bars), as well as technical replicates (hatched bars) and biological replicates (filled bars). From left to right, n = 1315, 1056, 1173, 1115, 865, 1253, 927, 1110, 1291, 1523, 1374, 1118, 850 and 988 cells per sample
