Fig. 3

Inhibition of HDAC5 enhances CB HSC homing by upregulating CXCR4 surface expression. a Fold change of mean fluorescence intensity of surface CXCR4 of human CB CD34⁺ cells after transfecting cells with indicated shRNA for individual HDACs. Individual shRNA knockdown efficiency was shown in Supplementary Fig. 2. Data pooled from three independent experiments are shown (n = 3, t-test, **p < 0.01). b Histogram of surface CXCR4 expression of human CB CD34⁺ cells treated with vehicle or HDAC5 inhibitor LMK235. Representative histogram from five independent experiments is shown. c Quantification of mean fluorescence intensity (MFI) of surface CXCR4 of human CB CD34⁺ cells treated with vehicle or HDAC5 inhibitor LMK235. None indicates the group without any treatment. Data pooled from five independent experiments are shown (n = 5, one-way ANOVA, ***p < 0.001). d Confocal imaging analysis of surface CXCR4 expression of human CB CD34⁺ cells treated with vehicle or LMK235. Green indicates CXCR4 expression; DAPI (blue) labels the cell nucleus. Representative images from two independent experiments are shown (the inset shows the amplified part of the image). Scale bar: 20 μm. e Human CB CD34⁺ cells migration toward human recombinant SDF-1, as quantified by flow cytometry. Data pooled from three independent experiments are shown (each dot represents an independent chemotaxis, n = 3 CB samples, one-way ANOVA, ***p < 0.001). f Migration of human phenotypic HSCs in CB CD34⁺ cells toward human recombinant SDF-1 (50 ng/mL), as quantified by flow cytometry. Data pooled from three independent experiments are shown (each dot represents an independent chemotaxis, n = 3 CB samples, t-test, **p < 0.01). g The percentage of human CD45⁺ cells in the BM and spleen of NSG mice 24 h after transplantation with 500,000 CB CD34⁺ cells that had been treated with vehicle or LMK235. Data pooled from seven independent experiments are shown (n = 7 mice per group, t-test, **p < 0.01, ***p < 0.001). h The percentage of human CD45⁺ cells in the BM of secondary recipient NSG mice that were transplanted with BM cells from primary mice used in the homing assay (n = 5 mice per group, t-test, *p < 0.05). For all panels, data are shown as dot plots (mean ± SEM)