Fig. 2: RECQ4 interacts and is required for MUS81 function in cells. | Nature Communications

Fig. 2: RECQ4 interacts and is required for MUS81 function in cells.

From: RECQ4-MUS81 interaction contributes to telomere maintenance with implications to Rothmund-Thomson syndrome

Fig. 2

A Immunoprecipitation (IP) was done from whole cell extracts expressing EGFP, EGFP-RECQ4 -WT, and the mutant EGFP-REQ4-Δ3. Proteins from each cell extract (500 µg) were incubated with GFP beads for 1 h and then resolved on SDS-PAGE, followed by western blotting detecting corresponding proteins. Input (IP), and FT fractions were run on different gels respectively. B Quantification of MUS81 binding from data represented in (A); n = 3 independent experiments; data are means ± SD; one-way ANOVA followed by Tukey’s multiple comparison test; p-value were adjusted by GraphPad Prism software; WT vs Δ3 **p = 0.0011. C Cell cycle-specific interaction between RECQ4 and MUS81. IP was done from whole cell extracts expressing EGFP-RECQ4-WT synchronised in different cell cycle phases. Protein from each extract (500 µg) was incubated with GFP beads for 1 h and then resolved on SDS-PAGE, followed by western blotting detecting corresponding proteins. D Graphical representation and representative images of binucleated cells by immunofluorescence with DAPI. Created in BioRender. Ashraf, R. (2025) https://BioRender.com/n17i536. Quantification of the average number of micronuclei per binucleated cell for each treatment; n = 3 independent experiments; data are means ± SD; one-way ANOVA followed by Tukey’s multiple comparison test; p-value were adjusted by GraphPad Prism software; siControl vs siRECQ4 **p = 0.0018; siControl vs siMUS81 ***p = 0.0003; siControl vs siMUS81/siRecq4 ***p = 0.0003. E Representative images and a graphical depiction of anaphase cells with bulky bridges. Created in BioRender. Ashraf, R. (2025) https://BioRender.com/n17i536. Quantification of the average number of bulky bridges per anaphase cell (DAPI positive bridges); n = 3 independent experiments; data are means ± SD; one-way ANOVA followed by Tukey’s multiple comparison test; p-value were adjusted by GraphPad Prism software; siControl vs siRECQ4 **p = 0.0015; siRecq4 vs WT/siRecq4 *p = 0.0155; WT/siRecq4 vs Δ3/siRECQ4 **p = 0.0025; WT/siRecq4 vs siMus81 *p = 0.0135; WT/siRecq4 vs siRecq4/siMus81 **p = 0.0094. Source data are provided as a Source data file.

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