Fig. 6: Evident aging phenotype in the Progerin-overexpressing assembloid model.

a PCA plot of the two first principal components on the gene expression value (FPKM) of all samples. Each sample represents data from 4 pooled assembloids from one batch. b Plot showing the log2 fold change of significant differentially expressed genes between Progerin-overexpressing assembloids (PROG_DOX) and control (WT_UNTR, WT_DOX, PROG_UNTR) samples. This list of genes was extracted after the comparison of the assembloid data with post mortem human brain data^32,33. c Western blot showing the protein levels of LAMIN B1 normalised to H3 housekeeping protein and batch corrected by normalising to the mean of the values for each batch. Outliers were calculated in GraphPad Prism using the ROUT method Q 1%. One-way ANOVA, with Tukey’s multiple comparison test was performed. For all conditions n = 4 with each point representing 3–4 pooled assembloids per batch, for 4 batches. Error bars represent mean ± SD. Data were plotted in GraphPad Prism 9.0.0. *p < 0.05, **p < 0.01, ***p < 0.001. d β-galactosidase staining for all the different assembloid conditions. Positive β-galactosidase areas were measured with ImageJ and normalised to the area of the section in each image and batch corrected by normalising to the mean of the values for each batch. Kruskal–Wallis test with Benjamini–Hochberg correction and Dunn’s multiple comparison test was performed. For all conditions n = 6 with each point representing one section per assembloid, per batch, for 3 batches. *p < 0.05, **p < 0.01, ***p < 0.001. Batch correction was applied by normalising each value to the mean of the values for each batch. Outlier removal was performed based on the Inter-Quartile Range (IQR) proximity rule. Data were plotted in R 4.2.2.