miR-221/222-sensitive genes in unperturbed HSC, MPP1, and MPP2 cells.

(A) Comparative differential expression analysis between miR-221/222-deficient and proficient HSC, (E) MPP1, and (F) MPP2 cells. The log2 fold-change expression values are plotted against the numbers of proficient cells expressing the gene. After calculating the linear regression curve, the 99% CI band (dashed red lines) determined the nondifferentially (gray dots) and differentially (red dots and gene symbols) expressed genes. Significantly higher expressed genes in miR-221/222-deficient cells are above the 99% CI limit and are expressed by more than 30% of all cells (green line). These genes are called “differentially expressed genes.” (B) Single-cell expression of Fos in HSC, MPP1, and MPP2 cells. The difference in the mean expression level of the indicated populations between proficient (open half violin) and deficient (filled half violin) cells are presented. (C) The RQ of Fos mRNA in bulk sorted miR-221/222-proficient (open bars) and miR-221/222-deficient (filled bar) HSC and MPP1 cells measured by RT-qPCR. RQ was calculated on the basis of Hprt expression in miR-221/222-proficient HSC. (D) The differential expression of AP-1 components Fos and Jun presented on UMAP of unperturbed miR-221/222-proficient and miR-221/222-deficient HSC, MPP1, and MPP2 cells. The numerical data for Fig 3A, 3B, and 3D–3F can be found in S2 Data, and the numerical data for Fig 3C can be found in S1 Data. AP-1, activator protein-1; CI, confidence interval; HSC, hematopoietic stem cell; MPP, multipotent progenitor; RQ, relative quantity; RT-qPCR, quantitative reverse transcription PCR; UMAP, Uniform Manifold Approximation and Projection for Dimension Reduction.