Extended Data Fig. 2: Phenotypes of MTHFD1 mutant alleles are suppressed by T-DNA insertion and CRISPR-Cas9 mutant alleles of THFS.
From: Photorespiration is linked to DNA methylation by formate as a one-carbon source
a, Images of 10-day-old mthfd1-1 CRISPR-Cas9 control (mT-cr23; no deletion in THFS detected by Sanger sequencing) and mthfd1-1 THFS knock out CRISPR-Cas9 (mtΔ-cr20; inversion between sgRNA111 and sgRNA243 detected by Sanger sequencing) T2 seedlings grown under LD. b, Agarose gel images of PCR genotyping for 8 T2 individuals per line (mT-cr23 and mtΔ-cr20). sgRNA target sites, genotyping primers, and expected amplicon sizes are indicated at right. c, Representative maximum projections of nuclear SDCpro-GFP expression acquired by CLSM z stacks from the ventral side of leaves of 3-week-old plants grown under LD. Scale bars, 50 μm. d, Corrected total cell fluorescence (CTCF) of SDCpro-GFP expression quantified from maximum projections. Box plots indicate medians (centre lines), IQR (boxes), and 1.5×IQR (whiskers). Indicated P value is from a two-sided Welch’s t-test (n = 10). e, Mean DNA methylation ratios over genes and TEs and the respective flanking regions in different sequence contexts. Two replicates (#1, #2) per genotype were analysed by whole genome bisulfite sequencing (WGBS). f, Representative pictures of 3-week-old mthfd1-3/mthfd1-3;THFS/thfs (mmTt) and mthfd1-3 thfs (mmtt) F3 plants grown under LD, leaf area quantification from automated phenotyping for mmTt (n = 4) and mmtt (n = 7), and relative DNA methylation levels in rosette leaves from mmTt, mmtt (n = 5) and Col-0 (n = 2) analysed by McrBC-qPCR at locus MG5. Data are presented as mean values +/- s.d.; indicated P value is from a two-sided Welch’s t-test.
