Fig. 7: HMGA2 regulates neuronal cell fate in an AT-hook 1-dependent manner.

a Domain organization and constructed mutants of HMGA2. b Analysis of droplet formation by recombinant PUB1_IDR-HMGA2 hook1 del (7.2 μM) in the presence or absence of mononucleosomes (800 nM) in a droplet formation solution and then observed with a DIC microscope. Scale bars, 10 µm. c Turbidity of incubation mixtures as in (b) was assessed by measurement of optical density at 400 nm (OD₄₀₀). Data are means ± s.d. (n = 3 independent experiments). d Immunohistofluorescence analysis of GFP and Ki67 in the neocortex of mice sacrificed (sac) at P1 after in utero electroporation (IUE) at E15.5 with an expression plasmid indicated in figure. Scale bar, 30 μm. e (left) Proportion of GFP-positive cells located in the cortical plate (CP), the intermediate zone (IMZ), or the ventricular zone (VZ)/subventricular zone (SVZ) in images as in (d). (right) Proportion of Ki67⁺ cells among GFP⁺ cells in images as in (d). Data are means ± s.d. (Localization: Control, HMGA2 full-length and HMGA2 hook1 del: n = 4, PUB1_IDR-HMGA2 hook1 del: n = 3, Ki67 staining: n = 3). Data are means ± s.d. Tukey’s multiple comparison test. f Proportion of Tbr2⁺, NeuroD1⁺, or Sox2⁺ cells among GFP⁺ cells detected by immunohistofluorescence analysis under the same conditions as in (d). Data are means ± s.d. (Control, HMGA2 hook1 del: n = 4, HMGA2 full-length: n = 4 (Tbr2), 6 (NeuroD1), 5 (Sox2)). Tukey’s multiple comparison test. g Expression of the indicated genes determined by RNA-seq from the neocortex at P1 after IUE at E15.5. (n = 5 independent experiments). h Enriched GO terms and their false discovery rate (FDR) values determined by functional annotation of genes whose expression was upregulated by overexpression of full-length HMGA2 in neocortical NPCs as measured by RNA-seq analysis. i Expression levels of all genes upregulated (left) or downregulated (right) by overexpression of full-length HMGA2 as determined by RNA-seq analysis. Data are presented as box plots, with the boxes representing the median and upper and lower quartiles and the whiskers indicating the range. Tukey’s multiple comparison test (two-sided).