Fig. 4: PTX3 is regulated by SH3RF3 and enhance BCSC properties.

a Expression heatmap of the genes differentially expressed in HMLER sublines and regulated by SH3RF3 overexpression. b PTX3 mRNA expression and its extracellular protein levels in conditioned media of various cancer cell lines after SH3RF3 overexpression and knockdown (n = 3 cell culturing). c Correlation of SH3RF3 and PTX3 mRNA levels in breast cancer cell lines (n = 51 cell lines). d Extracellular PTX3 levels in conditioned media of PTX3-overexpressing and control HMLE cells. e Tumorsphere formation of HMLE after PTX3 overexpression (n = 3 culturing experiments). f, g Flow cytometry analysis (f) and quantitation (g) of CD44⁺CD24⁻ CSC subpopulation in PTX3-overexpressing and control HMLE cells. Scale bar, 100 μm; n = 3 cell culturing experiments. h Tumor incidence in the mice injected with various numbers of HMLER cells at day 90. i GSEA analyses of CSC and MaSC-related gene sets in PTX3-overexpressing versus control cells. j GSEA analyses of Hedgehog and Hippo-regulated gene sets in PTX3-overexpressing versus control cells. k mRNA levels of Hedgehog and Hippo-regulated genes after PTX3 overexpression (n = 3 cell culturing experiments). Data represent mean ± SD. Statistical significance was determined by paired t-test (c) or two-tailed unpaired t-test (e, g, and k). The experiments in (b), (d), (e), (f), and (k) were repeated three times independently with similar results, and the data of one representative experiment are shown. Source data are provided as a Source data file.