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. 2021 Feb 19;22(3):e51989. doi: 10.15252/embr.202051989

Figure EV1. Establishing H3K27me3 mintbody to visualise H3K27me3 in living cells.

Figure EV1

  1. Amino acid sequence of the scFv region of H3K27me3‐mintbody derived from clone 2E12. Two amino acids that are substituted from the original 2E12 sequence are indicated with the regions of heavy and light chain variable fragments (VH and VL).
  2. Distribution of mintbody clones. Shown are single confocal sections of living mouse MC12 cells that transiently express mintbodies (sfGFP) from the original 2E12 and single (M86L) and double (M86L M158I) mutants. Scale bar = 10 μm.
  3. Model structures of 2E12 scFv. The 2E12 scFv structure was generated using ABodyBuilder (Leem et al, 2016) and the sites of substituted amino acid residues (coloured in orange) are indicated with the zoomed view. Substitutions from Met86 to Leu and Met158 to Ile can fill the gap (red arrows) and stabilise the hydrophobic cores in heavy (M86L) and light (M158I) chains. Nucleotide sequence of 2E12LI (M86L, M158I) is available under the accession number LC597262 in databases (DDBJ/EMBL/GenBank).