Fig. 8: BNC2-mediated regulation of ECM-related gene expression in mouse and human MF-HSCs.

A, B RT-qPCR data showing changes in gene expression upon BNC2 silencing in human (A) and mouse (B) MF-HSC cell lines and primary cells. Cells were transfected with BNC2-targeting siRNA (siBNC2) or a non-targeting control siRNA (siCTRL) for 48 h. Data for LX2 are from biologically independent experiments [n = 4 (COL1A1, COL3A1, COL5A1, LOXL1) or 5 (BNC2, COL1A2, COL5A2, LAMA5, ITGA1)] while data for human primary MF-HSCs were obtained using cells from different donors (n = 3). Data for EMS404 are from biologically independent experiments (n = 3 or 2 for Col5a1) while data for mouse primary MF-HSCs were obtained using cells from independent isolations [n = 3 (Col5a1, Col5a2, Lama5, Itga1, Loxl1) or 4 (Bnc2, Col1a1, Col1a2, Col3a1)]. Log₂ FC between siBNC2 and siCTRL-transfected cells are shown. C Simple western immunoassays used to monitor changes in BNC2 and type I collagen (COL-I) levels upon BNC2 silencing in LX2 (human) and EMS404 (mouse) cell lines. The lower panel shows the results of protein quantifications obtained from biologically independent experiments [(n = 3 (COL-I in EMS404), 4 (BNC2), or 5 (COL-I in LX2)]. BNC2 and COL-I levels were normalized to that of HSP90 or LMNA expression. Log₂ FC between siBNC2 and siCTRL-transfected cells are shown. MW, molecular weight markers. D Simple Western immunoassays used to monitor changes in BNC2 levels in LX2 cells treated with 1 μM pomalidomide (Pom.), thalidomide (Thal.), CC-885, Iberdomide (Iber.) or vehicle for 24 h. Lower panel shows the results of protein quantification obtained from three biologically independent experiments. Data were normalized to LMNA protein expression, and Log₂ FC between drug and vehicle-treated cells are shown. MW, molecular weight markers. E RT-qPCR data showing changes in gene expression upon treatment of LX2 cells as described for (D) (n = 3 biologically independent experiments or 2 for iberdomide). Log₂ FC between drug and vehicle-treated cells are shown. Pom., pomalidomide; Thal.; thalidomide; Iber., iberdomide. In all panels, bar graphs show means ± SD. Statistical significance was assessed using two-sided one-sample t test with Benjamini–Hochberg correction for multiple testing to determine if the mean log₂ FC was statistically different from 0. *P < 0.05, **P < 0.01, ***P < 0.001. Source data are provided as a Source Data file.