Fig. 6: Tethering of eIF4A1/2 to chromosomes accelerates chromatin decondensation.

A Time-lapse images of H2B-mPlum-FKBP HeLa cells expressing EGFP-FRB-eIF4A1 after adding 200 nM DMSO (control, - rapamycin) or rapamycin. Time is normalized to the first anaphase frame. Scale bars: 10 µm. B Time-dependent quantitation of the chromatin area of the experiments as in (A), normalized to the first anaphase frame. Dots represent mean ± s.e.m. from 12 (-rapamycin) and 14 (+ rapamycin) cells. C Same as in (A) but with the ATPase deficient mutant of eIF4A1 E183Q. D Same as in (B) but with the ATPase deficient mutant of eIF4A1 E183Q from 7 (-rapamycin) and 9 (+ rapamycin) cells. E Same as in (A) but with the RNA-binding mutant of eIF4A1 R362Q. F Same as in (B) but with the RNA-binding mutant of eIF4A1 R362Q from 9 (-rapamycin) and 10 (+ rapamycin) cells. G Same as in (A) but with EGFP-FRB-eIF4A2. H Same as in (B) but with EGFP-FRB-eIF4A2 with 15 (-rapamycin) and 17 (+ rapamycin) cells. I Same as in (A) but with EGFP-FRB-eIF4A3. J Same as in (B) but with EGFP-FRB-eIF4A3 with 10 (+ rapamycin) and 10 (-rapamycin) cells. K Time-dependent quantification of the chromatin area normalized to the first anaphase frame. HeLa cells stably expressing H2B-mCherry were co-transfected with GFP or eIF4A3-GFP and 40 nM control or a combination of eIF4A1 and eIF4A2 siRNA oligos for 48 h. Dots represent mean ± s.e.m. from each condition with 53 (EGFP + siCtrl), 37 (EGFP + sieIF4A1/2), 71 (eIF4A3-EGFP + siCtrl) or 45 (eIF4A3-EGFP + sieIF4A1/2) daughter chromatin masses. L Western blot showing the downregulation of eIF4A1/2 at 48 h post-transfection with 40 nM siRNA oligos and EGFP or eIF4A3-EGFP in HeLa cells stably expressing H2B-mCherry. Samples were analyzed with antibodies recognizing human eIF4A1/2. Actin serves as loading control. Source data are provided as a Source Data file.