Figure 2.

Effect of the Hit Compounds C6 and Its Analog C6-3 on HMBS in HepG2 Cells and Concentration-Dependent Surface Plasmon Resonance

(A and B) Western blotting and immunoquantification of the relative amount of HMBS in cell lysates from WT-HMBS stably transfected HepG2 cells treated with either C6 (A) or its analog C6-3 (B; see molecular structure in Table 3 and main text thereafter) at the indicated concentrations. DMSO (2%) was included in all samples. Representative blots are shown, and the histograms below represent the quantification of the relative HMBS levels (n = 3) using GAPDH as the protein loading control. Concentration-dependent binding of C6 (C) and C6-3 (D) to HMBS measured by SPR. The half-maximal binding (S_0.5) values were 83 ± 7 μM for C6, obtained by subsequent Octet measurements (C, inset), and 63 ± 3 μM for C6-3, obtained from the SPR binding isotherm (D). Data are presented as mean ± SD.