Fig. 7. The key amino acids mediating LLPS of LIN28A is required for its role in facilitating reprogramming of human fibroblast cells.

a Constructs used to investigate the function of mutations in IDRs of human LIN28A. b Reprogramming efficiency of NHDF cells transduced with human OCT4, SOX2, NANOG, and LIN28A variants in (A). c Number of iPSC colonies 21 days after OSNA transduction of NHDF cells. n = 3 biologically independent experiments. One-way ANOVA. Data are presented as mean values +/− SEM. d Representative STED immunofluorescence images of LIN28A and FBL in NHDF or iPSCs. FBL applied STED pattern. e FBL in NHDF and iPSC cells by STED. FBL exhibited a wreath-like structure in iPSCs. f NPM in NHDF and iPSC cells by STED. NPM exhibited a ring-like structure in iPSCs. g Statistical analysis of the FBL regularity in NHDF and iPSCs using Boyce-Clark semidiameter index. The larger the number, the more irregular the FBL. n = 15 nucleoli, unpaired two-tailed student’s t-test. h Immunofluorescence imaging showing OP-puromycin-labeled NHDF and iPSCs. Scale bar, 100 µm. i OP-Puro intensity statistical analysis of OP-puromycin-labeled NHDF and iPSC cells. n = 20 cells, unpaired two-tailed student’s t-test. The center line is the median, the bottom of the box is the 25th percentile boundary, the top of the box is the 75th percentile, and the top and bottom of the vertical line define the boundary of the data. Source data are provided as a Source Data file.