Fig. 7

CXCL2 is a regulatory target of QKI5 during monocytic differentiation. a–c Dual-uciferase reporter assays. Three independent experiments were performed, and data are presented as mean relative luciferase activity ± SD. a Diagram of QKI5 constructs and CXCL2 promoter constructs used. b The influence of QKI5/QKI5 M1 overexpression on activity of the wild-type (upper panel) and mutant CXCL2 promoters (lower panel). Left: Diagram of experiment. Right: 293T cells were co-transfected with the indicated combination of QKI5 and CXCL2 promoter constructs or control construct, then the activity of the promoter was calculated by the ratio of firefly and Renilla luciferase activities. Error bars indicate standard deviations of the three biological replicates. Asterisks indicate significant differences between the specified samples (*P value < 0.05, **P value< 0.01, t test). c The influence of QKI5 knockdown on activity of the wild-type CXCL2 promoter. Left panel: Diagram of experiment. Right panel: The same assay as above was conducted with the combination of shCtrl or QKI5 shRNA and CXCL2 promoter constructs or control construct. Asterisks indicate significant differences between the specified samples (*P value < 0.05). d Immuno-blot of CXCL2 and QKI5 in Ctrl- or QKI5/QKI5 M1-overexpressing (upper panel) and shCtrl- or shQKI5-treated (lower panel) THP-1 cells. e Experimental design of the two sets of rescue assays. f Percentage of CD14⁺/CD11b⁺ cells among THP-1 cells within shCtrl- or shCXCL2-treated populations, detected by flow cytometry in rescue 1, in which THP-1 cells were treated with shCtrl- or shCXCL2- shRNAs, followed by Ctrl or QKI5 /QKI5 M1 transduction. Average percentage of CD14⁺/CD11b⁺ cells is shown on the right. Error bars indicate standard deviations around the means of three biological replicates. Asterisks indicate significant differences between the specified samples (*P value < 0.05, **P value < 0.01, ns represents non-significant, t test). g Percentage of CD14⁺/CD11b⁺ cells among THP-1 cells within shCtrl- or shQKI5- treated population detected by flow cytometry in rescue 2, in which THP-1 cells were treated with shCtrl- or shQKI5- shRNAs, followed by Ctrl or CXCL2 transduction. Average percentage of CD14⁺/CD11b⁺ cells is shown on the right. h Schematic diagram of QKI5’s transcriptional regulatory function during monocytic differentiation