Fig. 5. The key amino acids mediating LLPS of LIN28A is required for its role in naive-to-primed pluripotency conversion of mouse ESCs.

a NANOG immunostaining of WT, Lin28a KO, and Lin28a KO mESCs transduced with full length WT LIN28A or Mut-LIN28A converted from the naïve state to the primed state. Scale bar, 10 µm. b Statistical analysis of NANOG protein fluorescence intensity of the above cells in (a). n = 64 cells. One-way ANOVA. c NANOG immunostaining of WT, Lin28a KO, and Lin28a KO mESCs transduced with full length WT LIN28A, single mutation variants, or IDR-fused variants, converted from the naïve state to the primed state. Scale bar, 200 µm. d Statistical analysis of NANOG protein fluorescence intensity of the above cells in (c). n = 64 cells. One-way ANOVA. e Representative STED immunofluorescence images of nucleoli in WT and Lin28a KO mESCs in the naïve state and the converted primed state. FBL applied STED pattern. f Typical FBL in WT and Lin28a KO mESCs in (e) by STED. Lin28a KO led to decreased size of FBL rings compared with WT when ESCs were converted to the primed state. Scale bar, 300 nm. g Statistical analysis of the number of FBL clusters in the above conditions. n = 10 nucleoli. Two-way ANOVA. h Statistical analysis of the relative FBL volume in the above conditions. n = 10 nucleoli. Two-way ANOVA. For (b, d) 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.