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. 2025 Dec 12;17:720. doi: 10.1038/s41467-025-67476-z

Fig. 5. RNAPII degradation strips chromatin-bound RNAPII and induces a global collapsing force and a local attractive force required for NSN-to-SN transition to take place.

Fig. 5

a Immunoblots of NSN, SN oocytes and HeLa cells. Black arrows mark phosphorylated (IIO), unphosphorylated (IIA) and tailless (IIB) RPB1. b Immunofluorescence images of NSN and SN nuclei. c Pie chart showing the distribution of pS2 CUT&Tag peaks in NSN oocytes. d Metagene plot of pS2 enrichment for genes of different expression levels in NSN oocytes. e Metagene plot of chromatin accessibility for genes of different expression levels in NSN oocytes and in SN oocytes. f Metagene plot of transcription for genes of different expression levels in NSN oocytes. g Still images from time-lapse movies of NSN oocytes treated with water or α-amanitin with tracking of chromocenters. h Automated quantification of MSD of chromocenters in NSN oocytes treated with water, 42B3-t21R, α-amanitin(α-aman.) or triptolide. Padjusted(42B3-t21R)=4.3521E-6; Padjusted(α-amanitin)=5.97982E-6; Padjusted(triptolide)=2.11141E-4. i Still images from time-lapse movies of NSN oocytes injected with water or overexpressing H2B-mClover3 with tracking of chromocenters. j Automated quantification of MSD of chromocenters in NSN oocytes injected with water or overexpressing H2B-mClover3. p = 0.0766. k Immunofluorescence images of α-amanitin-treated NSN oocytes overexpressing H2B-mClover3 or expressing αHIST-αLMN. Yellow arrowheads highlight the nucleolus. l, m Automated quantification of chromatin volume and surface area in α-amanitin-treated NSN oocytes overexpressing H2B-mClover3 or expressing αHIST-αLMN. Padjusted(H2B OE-chromatin volume)=5.42698E-4; Padjusted(αHIST-αLMN-chromatin volume)=0.00546; Padjusted(H2B OE-surface area)=0.89125; Padjusted(αHIST-αLMN-surface area)=5.01899E-5. n Immunofluorescence images of oil-injected NSN oocytes treated with water or α-amanitin. Yellow dotted circles highlight the oil droplet. o Immunofluorescence images of EU-low GV oocytes from wildtype and Npm2 cKO mice. The yellow arrowhead highlights the absence of a nucleolus. p Automated quantification of chromatin volume in GV oocytes from wildtype and Npm2 cKO mice. P(EU-high)=0.0924; P(EU-low)=0.6998. *p < 0.05. ****p < 0.0001. N.S., not significant. Box plots show median(middle line), mean(small squares), 25th and 75th percentiles(boxes), 5th and 95th percentiles(whiskers), and 1st and 99th percentiles(crosses). Unpaired, two-tailed Student’s t test (j, p), and one-way ANOVA followed by post-hoc Tukey’s multiple comparison test (h, l, m). The number of oocytes analyzed or loaded is specified in italics. Scale bars are 5 µm. Source data are provided as a Source Data file.