Fig. 2: ANLN and Pol II form condensates and colocalize at the gene TSS regions.

A Heatmap shows the CUT&Tag signals of ANLN and Pol II on the genome of KYSE150 cells. B Relative enrichment of CUT&Tag signals of ANLN and Pol II on TSS-gene body region. C Pie chart of ANLN and Pol II signals distribution in different regions at the genome. D Venn diagram of ANLN and Pol II peaks in KYSE150 cells. E Representative super-resolution images of nuclear ANLN (pink) and Pol II-pS5 (green) in HeLa cells. Scale bar, 3 µm. Cells were treated with or without 5% 1,6-HD for 60 s. Scale bar, 3 μm. F, G The number (F) and average area (G) of protein clusters per cell per focal plane. Clusters smaller than 12675 nm² were excluded based on previous studies. n = 57 (Control); n = 51 (1,6-HD). The error bars indicate the mean ± SDs. Box plots indicate the median (middle line, red), 25th and 75th percentiles (boxes), and minimum and maximum percentiles (whiskers). Two-tailed Student’s t tests were performed to determine the significance. H Relative fluorescence intensity of protein signals per cell per focal plane. n = 57 (Control); n = 51 (1,6-HD). The error bars indicate the mean ± SDs. Two-tailed Student’s t tests were performed to determine the significance. I, J Classify the identified clusters as type I (12675-25350 nm²), type II (25351-63375 nm²), and type III ( > 63375 nm²), showing the proportion of each cluster (n = 57). Scale bar, 3 µm. K Effect of 1,6-HD on the number of different types of clusters. n = 57 (Control); n = 51 (1,6-HD). The error bars indicate the mean ± SDs. Two-tailed Student’s t tests were performed to determine the significance. L Representative raw and reconstructed images of the interaction patterns between ANLN (pink) and Pol II-pS5 (green) large clusters (Type III). Scale bar, 3 µm. CUT&Tag analysis representative of two independent experiments, and other data are representative of at least three independent experiments with similar results. Source data are provided as a Source Data file.