Extended Data Fig. 7: Phosphorylation of METTL16 leads to dissociation with MRE11 by DNA damage-inducing reagents.
a, Immunoblots of METTL3 and METTL14 in different PDAC cell lines. β-Actin was used as a loading control. b,c, Relative METTL16 mRNA expression level (b) and the correlation of METTL16 mRNA and protein level (c) in the panel of PDAC cell lines. Data are presented as mean ± SD from three independent experiments (b). d, Quantification data for RPA32 foci formation in the indicated PDAC cell lines with or without 5 Gy X-ray irradiation treatment. Data are presented as mean ± SEM; each point represents a cell; the cells used for analysis in each experiment were from a single replicate; n indicates the cell number used for quantification in each group. e,f, HEK293T cells transfected with Flag-METTL16 vector were treated with the indicated DNA damage-induced agents. Cell lysates were subjected to immunoprecipitation with Flag beads and immunoblotted with the indicated antibodies. g,h, HEK293T cells transfected with Flag-METTL16 vector were treated with olaparib for the indicated time period or 10 Gy X-ray irradiation. Cell lysates were subjected to immunoprecipitation with Flag beads and immunoblotted with the indicated antibodies. i,j, Sanger sequencing results of METTL16 locus in SW1990 or PANC-1 cells with METTL16 WT or METTL16 knockout. Red arrows indicate the mutation sites. P-values are indicated (c,d). Statistical significance was determined by linear regression (c), or two-tailed unpaired t-tests (d). Experiments were repeated three times independently with similar results; data of one representative experiment are shown (a,e-h).
