Fig. 2

Phenotypic differences between HD and WT GMSLNs. a Representative microphotographs of 1) neurons differentiated from iPSHD22 cells, immunostained for microtubule-associated protein 2 (MAP2, red) and HTT (EM48 antibody, green), and counterstained with DAPI (blue); and 2) neurons differentiated from iPSHD22 cells, immunostained for HTT (EM48 antibody, green), and counterstained with DAPI (blue) following a 24 h incubation with or without 10 μM MG132. Scale bar, 20 μm. b Representative microphotograph of neurons differentiated from iPSHD22 cells acquired via TEM. Micrographs show high lysosomal and autophagosomal content, mitophagy, and nuclear envelope indentation. c Representative FC analysis of live neurons stained with LysoTracker Green; HD (green), WT (blue), and without staining (grey). The bar plot demonstrates median fluorescence intensity from three independent experiments. d Representative microphotographs of terminally differentiated WT and HD neurons stained with DAPI demonstrating nuclear indentations. Scale bar, 50 μm. The bar plot demonstrates morphometric quantification of nuclear irregularity index using 786–1340 nuclei per data point. e The bar plot demonstrates morphometric quantification of nuclear irregularity index in cultures of iPSHD22 derived neurons 4 days after transfection with antisense oligonucleotides: LNA(T) – specifically knockdown mHTT, LNA(S) – scrambled oligonucleotide, Control – no transfection. 754–1405 nuclei were count per data point. f The bar plot demonstrates morphometric quantification of nuclear irregularity index in cultures of WT PSCs (endo-iPS12, IPSRG2L and hESM01) derived neurons 9 days after infection with lentiviral constructs HTTQ15-1exon and HTTQ138-1exon. 754–1405 nuclei were count per data point. g Bar plot of the average mean nuclear irregularity index of HD (iPSHD11, iPSHD22, and iPSHD34) and WT (endo-iPS12, IPSRG2L, and hESM01) neurons after treatment with the indicated drugs. Morphometric quantification was conducted by using 2785–4649 nuclei per data point