FIG 5.

NeoB reduced lipid accumulation in hepatocyte-derived cell lines. (A) Oil red O staining was performed as described in Materials and Methods with Huh7.5.1 and Huh7-25 cells treated with 0.2% DMSO (control; a and e), 20 μM NeoB (b and f), 20 μM NeoA (c and g), or 30 μM 5CPPSS-50 (d and h) for 72 h. (B) mRNAs for SREBP1c and GAPDH were detected in Huh-7.5.1 cells treated with or without the indicated compounds in the presence of 5 μM TO-901317 for 24 h. (C) Uncleaved and cleaved forms of SREBP1 and actin proteins were detected by immunoblotting using Huh-7 cells treated with or without NeoB (5, 10, and 20 μM) as described in Materials and Methods. The arrow indicates the cleaved SREBP1 protein. The values below the panels indicate the band intensities of cleaved SREBP1 and actin proteins. (D and E) Oil red O staining was performed on HCV-infected Huh7 cells or primary human hepatocytes treated with 0.2% DMSO (control; i, i-2, and l), 20 μM NeoB (j, j-2, and m), or 20 μM NeoA (k, k-2, and n) for 72 h. Panels i-2, j-2, and k-2 are higher-magnification images of the boxed areas shown in panels i, j, and k, respectively.