Fig. 1

Characterizing the genomic stability and differentiation consistency of the temperature sensitive SN4741 cell line. A A representative karyogram of SN4741 cells, indicating structural instability (M; marker chromosomes) and unstable triploidy. B A stacked bar plot summarizing the aneuploidy frequency of each chromosome over 20 SN4741 karyotypes. C Assaying expression of dopaminergic neuron markers by RT-qPCR indicates that Foxa2, Nr4a2,Slc6a3, and Th remain at similar expression levels when SN4741 cells are shifted from the permissive temperature (37 °C) to the non-permissive temperature (39 °C). D UMAP plot of scRNA-seq at the permissive and non-permissive temperatures indicates that cells at each temperature are transcriptionally distinct. E Analysis of scRNA-seq data demonstrates that shifting the cells to the non-permissive temperature is accompanied by a shift in cell cycle stage from G2M and S phases to primarily G1 phase. F Violin plots generated with scRNA-seq data show that Mki67, a marker of cellular proliferation, and Nes, a neural stem cell marker, are both expressed at the permissive temperature (37 °C), with little to no expression at the non-permissive temperature (39 °C). G Violin plots generated with scRNA-seq data show that transcripts associated with immature neurons are upregulated when SN4741 cells are shifted to the non-permissive temperature. H Violin plots generated with scRNA-seq data show that expression of DA neural markers, Aldh1a1, Foxa2, Lmx1b, Nr4a2, Pitx3, Slc6a3, and Th, remain at similar levels when SN4741 cells are shifted to the non-permissive temperature. (NS = Not significant, * = p < 0.05, ** = p < 0.01, *** = p < 0.001, **** = p < 0.0001)