Journal: Nucleic Acids Research

Article Title: Enhanced or reversible RNA N6-methyladenosine editing by red/far-red light induction

doi: 10.1093/nar/gkaf181

Figure Lengend Snippet: Dynamic m 6 A regulation modulates hESC endoderm differentiation. ( A ) MeRIP-RT-qPCR of the relative m 6 A levels at the SOX2 A1398 site in hESCs transduced with the light-induced editors. The cells were reversibly edited by the far-red light (740 nm, 2 Day) or far-red/red light (740 nm, 2 Day; 660 nm,1 Day) exposure. ( B ) SELECT of the m 6 A at the SOX2 A1398 in hESCs upon the conditions. A lower C t value (i.e. a higher amount of the full-length SELECT product) indicated a lower level of m 6 A at the probed A1398 site. ( C ) The relative mRNA abundance of NANOG, SOX2, OCT4, SOX17, CXCR4 ,and FOXA2 in hESCs under different light exposure conditions. ( D ) Flow cytometric analysis of endoderm differentiation efficiency of hESCs by surface marker CXCR4. All results were calculated by normalizing data from each sample to that from the condition of ΔphyA-dCas13b plus sgRNA-22. Values and error bars reflect the mean, S.E.M. of three independent biological replicates. Source data are provided in the source data file.

Article Snippet: The H9 human embryonic stem cell (hESC) line was cultured in mTeSR1 medium (STEMCELL Technologies, cat. no. AB217641) on Matrigel-coated plates (Corning, cat. no. 354277) with daily medium changes.

Techniques: Quantitative RT-PCR, Transduction, Marker