Figure 4
Pharmacological inhibition of BMP1 decreases the proteolytic cleavage of LDLR and increases the cellular uptake of LDL. (A) Immunoblot analysis of LDLR in cell lysates and conditioned media from HepG2 cells treated with inhibitor UK383367 (10 μM) for 18 h. Blot images were cropped to highlight the FL and NTF bands in cell lysates and conditioned media, respectively, using the Ab14056 antibody. Full blot images are presented in the Supplementary western blot dataset. (B) Densitometric analysis of the NTF from (A). Data shown as mean ± SEM, ***p < 0.001, n = 3. (C) BODIPY®-LDL uptake in HepG2 cells treated with or without inhibitor UK383367 (10 μM) for 6 h. Data expressed as percentage of control and shown as mean ± SEM, statistical analysis performed using a Student’s T-test **p < 0.01, n = 3. (D) Representative data from flow cytometry showing the change in cell surface fluorescence of LDLR between HepG2 cells treated with inhibitor UK383367 (10 μM) for 18 h (blue line) and untreated cells (red line); primary antibody (Ab14056) only (grey line). (E) Quantitation of the cell surface expression of LDLR in HepG2 cells treated with or without UK383367 as analysed by flow cytometry in (D). Data expressed as percentage of control and shown as mean ± SEM, statistical analysis performed using a Student’s T-test **p < 0.01, n = 3.
