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. 2019 Nov 4;10(11):830. doi: 10.1038/s41419-019-2054-7

Fig. 4. Downregulation of ASPP2 promotes mevalonate metabolism by activation of SREBP-2.

Fig. 4

a Immunofluorescent images of HCC-LM3 cells showing the cellular location of endogenous ASPP2 and SREBP-2. ASPP2 and SREPB-2 were stained green and red, respectively. DAPI (blue stain) was used to stain the nucleas.Scale bars: 40 μm. b Western blot of the immunoprecipitated ASPP2-SREBP-2 complex in total cell lysates (Left) or nuclear proteins lysates (Right) with anti-ASPP2 in HCC-LM3 cells. c Western blots of V5-tagged ASPP2 co-transfected with HA-SREBP-2 into HEK293T following IP with anti-V5 antibody in total cell lysates (Left) and nuclear proteins lysates (Right). d Western blots of HA-tagged SREBP-2 co-transfected with ASPP2-V5 into HEK293T following IP with anti-HA antibody in total cell lysates (Left) and nuclear proteins lysates (Right). e Luciferase activity of Hamster HMGCS1, human LDLR promoters and three Sterol regulatory elements (3SRE) with respect to reporter vector pGL3.0. The constructs were co-transfected with an internal control pRL-TK vector into HCC-LM3 cells with silencing of ASPP2 (left) and Huh-7 cell with ASPP2 overexpression (right). f The number of spheres per 1000 HCC cells with siRNA negative control or siRNA Srebp-2 after knock down of ASPP2. g Western blot of HMGCR and HMGCS1 protein in HCC-LM3 cells infected with LV-shASPP2 or LV-shNon and treated with siAspp2 and/or siSrebp2. Asterisk (*) indicates P < 0.05, asterisks (**) indicates P < 0.01