Supplementary Figure 4: Nuclear pores and INMPs recruit STUbL/RENi to the nuclear periphery, and do not prevent aberrant recombination in heterochromatin.

(a) Quantitation of the experiment described in Fig. 4a shows the frequency of cells with a ‘clustering’ phenotype after Lamin RNAi, relative to cells with intact Lamin signal (normal) and cells without Lamin (no Lamin). (b) IF analysis of cells expressing GFP-Spag4, fixed after Lamin RNAi and stained for Nup153 (nuclear pore marker), Koi and GFP. (c) Western analyses of cells expressing GFP-Dgrn and FHA-dRad60 show the levels of Dgrn (left) or dRad60 (right) proteins after Nup107+Koi+Spag4 RNAi relative to Ctrl RNAi. Actin and tubulin are loading controls. (d) IF analysis of cells expressing FHA-dRad60 and GFP-Dgrn with anti-GFP (top) or-HA (bottom) antibodies, after RNAi depletion of the indicated proteins. (e) IF analysis of cells expressing FHA-dRad60 and GFP-Dgrn from the experiment described in Fig. 4c were stained for Nup153, Koi, and GFP (left) or HA (right). (f) Cells were fixed 60 min post IR after RNAi depletions, and analyzed by IF to detect Rad51 foci. Quantifications show the frequencies of foci relative to DAPI-bright and DAPI-weak regions (p < 0.0001 for Smc5/6 RNAi versus Ctrl RNAi in DAPI-bright, n > 160 cells; error bars represent mean ± s.d. from three independent experiments, whereas the sample size is the total number of cells for each RNAi, pooled across the three experiments). (g) Quantitation of DNA filaments connecting dividing cells in cells fixed and processed as in Fig. 1f (n > 700 cells, error bars: mean ± s.d. from three independent experiments). Smc5 and Ctrl RNAi samples are the same as in Fig. 1f. (h) Quantitation of foci in DAPI-weak for the experiment described in Fig. 4d. Ctrl/C = control. Scale bars = 1 μm. Images are middle Z-stacks of nuclei. Exact n values are in Supplementary Table 1.