Fig. 1: Cell viability effects of the genome-wide deposition of m6dA by expression of bacterial N6-MTases. | Communications Biology

Fig. 1: Cell viability effects of the genome-wide deposition of m6dA by expression of bacterial N6-MTases.

From: Genome-wide deposition of 6-methyladenine in human DNA reduces the viability of HEK293 cells and directly influences gene expression

Fig. 1

a Design of the N6-MTase expression constructs which were stably integrated into the genome of HEK293 cells. The reverse tetracycline-controlled transactivator 3 (rtTA3) binds the TRE3G promoter in the presence of dox, thus activating the expression of the N6-MTase. A fluorophore is co-expressed over an internal ribosomal entry site (IRES). b Schematic depiction of the experimental workflow used for the competitive proliferation assay of N6-MTase versus HEK293 cells. c, d Flow cytometry results of the competitive proliferation assay for EcoDam (WT and D181A mutant), or CcrM (WT and D39A mutant), against HEK293 cells. The fraction of GFP-positive (GFP+) cells relative to non-fluorescent cells was calculated and normalized to day 4 of dox treatment. e Schematic depiction of the experimental workflow used for the competitive proliferation assay of N6-MTase 1 (GFP+) versus N6-MTase 2 (dsRed+). f, g Flow cytometry results of the competitive proliferation assay for EcoDam or CcrM variants (GFP+) versus the corresponding inactive mutants (dsRed+). The fraction of GFP+ cells was calculated and normalized to day 4 of dox treatment.

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