Figure legends
Figure 1. Increased single-cell transcriptome heterogeneity of suspension-cultured CHO cells over time. Single-cell transcriptome sequencing was performed on 85, 70, and 78 cells from cultures maintained for 4, 8, and 11 days, respectively. Principal components 1 (proportion of variance, 0.31) and 2 (proportion of variance, 0.077) are plotted. Blue, black, and red plots represent individual day 4, day 8, and day 11 cells, respectively. Variances in principal component 1 between the different culture times had a p-value < 2.2e-16.
Figure 2. Single-cell resolution mitochondrial genomic heterogeneity in suspension-cultured CHO cells. Genotypes of 498 variable loci in each cell were visualized.
Figure 3. Correlation between mutation rate and culture time. Single-cell resolution genomic mutation analysis was performed on the mitochondrial genomes of CHO cells cultured for different periods. No statistical correlation was observed between culture time (Days) and the heteroplasmic variant rate (rate of new mutations at called variable loci; p = 0.14).
Figure 4. Cell cycle–dependent subpopulations in traditional adherent CHO cultures. Single-cell transcriptome sequencing was performed on 24 G1 cells (circles) and 8 G2 cells (crosses).
Figure 5. Hidden subpopulations in suspension cultures of CHO K1 cells. Individual D4 cells clustered into groups with high enolase expression (2000–4200 RPM, red) and low enolase expression (0–2000 RPM, black). (a) Histogram of enolase expression degree (RPM) in suspension CHO K1 cells after 4 days. (b) A colored plot of principal components 1 and 2 in Day 4 suspension cultures of CHO K1 cells.