Journal: Cell Death & Disease

Article Title: IL-6-C/EBPβ signaling drives monocytic differentiation of murine cultured lymphoid progenitors with immunoregulatory properties

doi: 10.1038/s41419-025-07930-4

Figure Lengend Snippet: a Scatter plot of DEG (FDR < 0.05, >2-fold or <0.5- fold) between undifferentiated cCLPs and cCLPs after 24 h incubation with FL/SCF/IL-6 (D1). b Venn diagram of genes up-regulated in D1 to cCLPs with enrichment analysis of “Cell Differentiation” by GO Biological process and those up-regulated in D3 to cCLPs. c Heatmap view of 57 genes overlapped in ( b ). Color code values indicate the Z-score for each gene across samples. d Gene expression of Cebpb ( left panel ) and Id1 ( right panel ) in indicated samples. The values are shown as log 2 (CPM + 4) from RNA-seq data. e Representative FACS plots of cCLP-Ms differentiated from single cCLP clones where indicated genes were targeted by Crispr/Cas9. The name of a representative clone targeted to each gene is shown (＃2–3, ＃2-17, #2-16). f Frequency of wells with more than 100 live cells in ( e ). g Frequency of wells with less than 40% CD11b + CD115 + cells. h Representative histogram of GFP expression ( left panels) , plots for CD11b and CD115 expression on GFP - gate ( middle panels ), or GFP + gate ( right panels ) in cCLPs on day 4 after retroviral transduction with pMIG empty vector ( upper panels ) or pMIG/ C/EBPβ ( lower panels ). i Percentages of CD11b + CD115 + cells in the indicated fractions in ( h ). Data are mean ± SD with statistical significance determined by one-way ANOVA (in d ) or two-way ANOVA (in i ) with multiple comparisons. The p values are represented as **, <0.01; ***, <0.001; ****, <0.0001. n.s. not significant.

Article Snippet: The amplified PCR product was inserted at the Xho I site of the pMSCV-IRES-GFP retroviral vector (pMIG) using NEBuilder HiFi DNA Assembly Master Mix (New England Biolabs, E2621S).

Techniques: Incubation, Cell Differentiation, Gene Expression, RNA Sequencing, Clone Assay, CRISPR, Expressing, Retroviral, Transduction, Plasmid Preparation