Figure 4.
Single-cell transcription and GSEA profiles of LK and MPP cells from Sptlc1+/+and Sptlc1−/−. (A) A total of 3140 Sptlc1+/+ and 2808 Sptlc1−/− cells identified as LK by flow cytometry were divided into clusters based on individual gene-expression profiles. The plot shows a 2-dimensional representation (t-SNE) of transcriptome profiles of the Sptlc1+/+ and Sptlc1−/−, with each cluster highlighted in a different color. The integrated Sptlc1+/+ and Sptlc1−/− cells were represented in t-SNE plots. (B) The percentage of cells expressing relative lineage-specific markers in individual clusters from Sptlc1+/+ and Sptlc1−/− LK populations. (C) Heat maps of Sptlc1+/+ and Sptlc1−/− LK populations expressing myeloid-specific gene markers (Mpo, Elane, Ctsg, Prtn3, Irf8, and Cebpe) and erythroid-specific gene markers (Apoe, Klf1, Rhd, Hba.a1, Gata1, Gata2, Car1, and Car2). (D) Violin plots of the distributions of expression of Mpo, marker of the myeloid population, in LK cells. Comparison of expression levels for LK cells in clusters C1, C3, C4, C5, C7, and C9 of Sptlc1+/+ and Sptlc1−/− is shown. (E) Violin plots of the distributions of expression of Car2, a marker of the erythroid population. Comparison of expression levels for LK cells in clusters C2 and C5 of Sptlc1+/+ and Sptlc1−/− is shown. (F) The percentage of myeloid, erythroid, and multilineage cells in the Sptlc1+/+ and Sptlc1−/− LK populations. (G) A total of 3099 Sptlc1+/+ and 1997 Sptlc1−/− cells identified as MPPs by flow cytometry were divided into clusters based on individual gene-expression profiles. The plot shows a 2-dimensional representation (t-SNE) of transcriptome profiles of the Sptlc1+/+ and Sptlc1−/−, with each cluster highlighted in a different color. The integrated Sptlc1+/+ and Sptlc1−/− data set was represented in a t-SNE plot. (H) Violin plots of the distributions of expression of Hspa5 and Pdia6, indicators of ER stress, in Sptlc1+/+ and Sptlc1−/− MPP cells. (I-J) GSEA analysis of RNA-seq data showing indicated gene sets with differentially regulated genes between Sptlc1+/+ and Sptlc1−/− in LK (I) and MPP (J) cell populations, respectively. Heat map of top 30 genes were displayed. Relative expression levels are indicated. FDR, false discovery rate; NES, normalized enrichment score.

Single-cell transcription and GSEA profiles of LK and MPP cells from Sptlc1+/+and Sptlc1−/−. (A) A total of 3140 Sptlc1+/+ and 2808 Sptlc1−/− cells identified as LK by flow cytometry were divided into clusters based on individual gene-expression profiles. The plot shows a 2-dimensional representation (t-SNE) of transcriptome profiles of the Sptlc1+/+ and Sptlc1−/−, with each cluster highlighted in a different color. The integrated Sptlc1+/+ and Sptlc1−/− cells were represented in t-SNE plots. (B) The percentage of cells expressing relative lineage-specific markers in individual clusters from Sptlc1+/+ and Sptlc1−/− LK populations. (C) Heat maps of Sptlc1+/+ and Sptlc1−/− LK populations expressing myeloid-specific gene markers (Mpo, Elane, Ctsg, Prtn3, Irf8, and Cebpe) and erythroid-specific gene markers (Apoe, Klf1, Rhd, Hba.a1, Gata1, Gata2, Car1, and Car2). (D) Violin plots of the distributions of expression of Mpo, marker of the myeloid population, in LK cells. Comparison of expression levels for LK cells in clusters C1, C3, C4, C5, C7, and C9 of Sptlc1+/+ and Sptlc1−/− is shown. (E) Violin plots of the distributions of expression of Car2, a marker of the erythroid population. Comparison of expression levels for LK cells in clusters C2 and C5 of Sptlc1+/+ and Sptlc1−/− is shown. (F) The percentage of myeloid, erythroid, and multilineage cells in the Sptlc1+/+ and Sptlc1−/− LK populations. (G) A total of 3099 Sptlc1+/+ and 1997 Sptlc1−/− cells identified as MPPs by flow cytometry were divided into clusters based on individual gene-expression profiles. The plot shows a 2-dimensional representation (t-SNE) of transcriptome profiles of the Sptlc1+/+ and Sptlc1−/−, with each cluster highlighted in a different color. The integrated Sptlc1+/+ and Sptlc1−/− data set was represented in a t-SNE plot. (H) Violin plots of the distributions of expression of Hspa5 and Pdia6, indicators of ER stress, in Sptlc1+/+ and Sptlc1−/− MPP cells. (I-J) GSEA analysis of RNA-seq data showing indicated gene sets with differentially regulated genes between Sptlc1+/+ and Sptlc1−/− in LK (I) and MPP (J) cell populations, respectively. Heat map of top 30 genes were displayed. Relative expression levels are indicated. FDR, false discovery rate; NES, normalized enrichment score.

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