Additional file 1 of Spliceosome inhibitor induces human hematopoietic progenitor cell reprogramming toward stemness
Creators: name: Dong, Liaoliao
name: Wei, Chuijin
name: Xiong, Shumin
name: Yu, Ping
name: Zhou, Ren
name: Cheng, Lin
Publisher: name: figshare
Year: 2022
Subjects: subject: Genetics
subject: FOS: Biological sciences

description: Additional file 1: Figure S1. Expression levels of splicing factors increased along with HSC expansion. A Representative FACS plot(left) and pie diagram(right) showed the percentage of HSPC subpopulations of human UCB HSCs after 7-day-cultured in vitro. Cell surface markers used to gate cell populations were listed. B Folds change of cell quantity compared with that of initial HSCs after 7-day-cultured in vitro. C Heatmap of splicing factors expression levels in HSCs, Multipotent Progenitors and Committed Progenitors. Many splicing factors gradually increased along with HSC differentiation. D Percentage variation of HSPC among samples treated with different PlaB concentrations, supplement to figure 1A. Data represented as means ± SDs from N = 3 duplicates. 2-tailed unpaired t test with unequal variance; n.s.: p &gt; 0.05, *p &lt; 0.05, **p &lt; 0.01, ***p &lt; 0.001. Figure S2. Basic information of sc-RNAseq. A, B Distribution of confidently mapped reads information on scRNA-seq. scRNA-seq of the cells at both timepoints before and after PlaB treatment included 11,048 and 12,173 individual cells together with 2933 and 3054 median genes and 37,771 and 29,461 mean confidently mapped reads per cell. C UMAP visualization based on 10× scRNA-seq before and after PlaB treatment. MD cells: Monocytes and Dendritic cells, EBM cells: Eosinophils, Basophils and Mast cells. Figure S3. Basic information of CellTagging and lineage tracing. A The CellTagging workflow: a lentiviral construct contains a heritable 8-bp random CellTag barcode in the 3’ UTR of GFP, followed by an SV40 polyadenylation signal. Transduced cells express unique CellTags, enabling tracking of clonally related cells. B Number of CellTags detected in scRNA-seq samples. The number of celltag inserted in each cell was from 1 to 6 and the average number was 1. C Number of paired and individual CellTags detected in scRNA-seq samples before and after PlaB treatment. 7888 cells before treatment and 9570 cells after that have been detected with celltags and 6290 cells between two timepoints were inserted with the same celltag. D Topological map of cells tracing by CellTagging, related with Figure 2D. Specific genes of each cluster were shown in PAGA layout. Figure S4. Basic information of mitochondrial DNA mutation and lineage tracing. A The principle of lineage tracing by mitochondrial DNA mutation. Each cell has multiple mitochondria, which in turn contain many copies of mtDNA that may acquire somatic mutations over time. Correlation analysis of mutation patterns enable tracking of clonally related cells. B Based on the scRNA-seq data, distribution of coverage of the mitochondrial genome, numbers of mutations, frequency of mutations and numbers of mutations per cell were calculated. C Hierarchical clustering of mitochondrial genotyping profiles (rows) for cells on two timepoints. D Topological map of cells tracing by mitochondrial mutation, related with Figure 2D. Specific genes of each cluster were shown in PAGA layout.