Extended Data Fig. 4: Analysis of the influence of dCas13 binding on RNA stability, distribution, and translation, related to Fig. 1j.
From: Programmable control of spatial transcriptome in live cells and neurons
a-f, Representative microscopic images showing dCas13 expression and RNA FISH signals of endogenous GAPDH mRNA (a), human ACTB mRNA (b), mouse Actb mRNA (c), NDUFS2 mRNA (d), SDHC mRNA (e), and SDHD mRNA (f) in HeLa cells (a,b,d,e,f) or Neuro-2a cells (c) expressing dCas13 and the corresponding targeting gRNAs compared with gNT. Calibration bar indicates the fluorescence intensity of RNA FISH signals. Scale bar, 50 µm. g-l, Violin plots quantifying the integrated intensity of RNA FISH signals of the target mRNA in individual cells expressing dCas13 and the corresponding targeting gRNAs compared with gNT. n, cell number. m-r, Violin plots quantifying the Polarization Index and Dispersion Index of the target mRNA in individual cells expressing dCas13 and the corresponding targeting gRNAs compared with gNT. n, cell number. s and v-x, Violin plots quantifying the integrated intensity of IF signals of proteins translated from the target mRNA in single HeLa cells expressing dCas13 and the corresponding targeting gRNAs compared with gNT. n, cell number. t-u, Column plots showing the mean fluorescence intensity of human ACTB protein (t) and mouse ACTB protein (u) IF signals analyzed by flow cytometry in HeLa cells (t) or Neuro-2a cells (u) expressing dCas13 and the corresponding targeting gRNAs compared with gNT. Each dot represents one biological replicate. Data presented as means ± standard deviation. n = 3 wells per group. All the P values were calculated using two-sided, unpaired Student’s t-test between gT and gNT groups.
