Extended Data Fig. 8: Validation of reverse translocation (γ) of cluster 4 genes in G1 by smFISH-HCR and subcellular RNA kinetics in the context of m⁶A post-transcriptional modification.

a, schematics of HeLa-FUCCI cells. The cell line expresses two fluorescently-tagged cell cycle marker proteins (red, RFP-CDT1 and green, GFP-Geminin); these two markers have co-expression during G1-S transition (yellow). b, after plotting the normalized intensities of GFP and RFP, we obtained G1 and G2/M phases based on the thresholds indicated in the plots. c, boxplots comparing the DR values of ACTB and MKI67 in G1 and G2/M based on the cell label in c. d, Pie chart describing m⁶A-RNA methylation in the gene pool (see Methods). e, Violin plots showing the difference of synthesis (left) and degradation rates (right) between m⁶A-RNA and non-m⁶A-RNA from scEU-seq dataset. Paired t-test. Boxplots indicate mean and 25-75% quantile. f, Boxplots comparing the four parameters estimated for m⁶A (n = 476 genes) and non-m⁶A RNAs (n = 89 genes) across three cell-cycle phases. Units of α is RNA concentration/hr, where RNA concentration is defined by copies/voxel (voxel = 200 nm x 200 nm x 350 nm). Units of β, λ, γ are hr⁻¹. ** p < 0.01, Wilcoxon test. Data shown as means (notches), 25-75% quartiles (boxes) and ranges (vertical lines).