fatostatin  (MedChemExpress)

Journal: Cell reports

Article Title: SREBP-1 upregulates SOAT1 to promote tumor growth by preventing lipotoxicity

doi: 10.1016/j.celrep.2025.116896

Figure Lengend Snippet: (A) Schematic of the process of SREBP-1 activation and its potential role in regulating SOAT1 expression. The inhibitor fatostatin blocks trafficking of the SCAP/SREBP-1 complex, and PF429242 suppresses SREBP-1 cleavage by inhibiting site 1 protease (S1P). The diagram was created using BioRender. (B–G) Western blot analysis of the indicated proteins (B, C, and F) and representative fluorescence images of LDs stained with BODIPY 493/503 (D, E, and G) in GBM or lung cancer cells following treatment with fatostain (Fato) (B and D) or PF429242 (C and E) for 24 h or SREBP-1 knockdown using lentivirus-mediated shRNA for 48 h (F and G). (H) Schematic of the putative SREBP-1 binding sites (SREs) and a negative control (NC) region in the SOAT1 promoter (top). Arrows indicate the positions of primers used for PCR analysis following chromatin immunoprecipitation (ChIP) with either control immunoglobulin G (IgG) or anti-SREBP-1 antibody (bottom). (I) Measuring the activities of the SOAT1 promoters using a pGL3-luciferase (Luc) reporter plasmid in HEK293-FT cells in response to the expression of N -terminal SREBP-1a or -1c isoforms for 48 h (J) Mutagenesis of the putative SRE in SOAT1 promoter and measuring its activity in response to expression of N-terminal SREBP-1a or -1c isoforms for 48 h (K) Real-time PCR analysis of SOAT1 gene expression in GBM30 or H1299 cells following adenoviral expression of FLAG-tagged N-terminal SREBP-1a (Ad- 1aN ), or SREBP-1c (Ad- 1cN ) isoforms compared to a control vector (Ad- null ) for 48 h (L and M) Western blot (L) and fluorescence imaging of LDs stained with BODIPY 493/503 (M) in GBM30 and H1299 cells after adenoviral expression of FLAG-tagged N-terminal SREBP-1a or -1c isoforms for 48 h under 5% FBS conditions. All experiments were performed in three independent biological replicates. For (D), (E), (G)–(K), and (M), data are presented as means ± SD. Nuclei were counterstained with Hoechst 33342 (blue). Scale bar, 10 μm. Quantification of LDs was performed using ImageJ from >30 cells, for (D), (E), (G), and (M). Statistical significance was analyzed by one-way ANOVA compared with vehicle-treated controls for (G) or Student’s t test for (D), (E), (H), (I)–(K), and (M). See also .

Article Snippet: Fatostatin , ChemBridge Corporation , Cat# 5533803.

Techniques: Activation Assay, Expressing, Western Blot, Fluorescence, Staining, Knockdown, shRNA, Binding Assay, Negative Control, Chromatin Immunoprecipitation, Control, Luciferase, Plasmid Preparation, Mutagenesis, Activity Assay, Real-time Polymerase Chain Reaction, Gene Expression, Imaging

Journal: Nature Communications

Article Title: The UFL1-AKT positive feedback loop promotes breast cancer progression by enhancing lipid synthesis

doi: 10.1038/s41467-026-68492-3

Figure Lengend Snippet: A – D AKT1 UFMylation and phosphorylation was determined by IP and IB assays in MDA-MB-231 and MDA-MB-468 cells treated with 20 μM DKM 2-93 (UFMylation inhibitor), 6 μM MK-2206 (AKT inhibitor), or a combination of these inhibitors for 24 h. E–H Representative images of ORO staining ( E ) and BODIPY staining ( G ) in the indicated breast cancer cell lines treated with 1 mM sodium citrate for 12 h, followed by quantification of lipid content in these cells ( F , H ). Scale bars, 60 μm in ( E ), 25 μm in ( G ). n = 3 independent experiments per group. I–K , M Growth curves ( I ), tumor weights ( J ), representative images of ORO staining ( K ), and lipid content quantification ( M ) of the indicated xenograft tumors. Scale bars, 60 μm. n = 5 mice per group. The drug was administered every three days until the mice were euthanized. The arrow indicates the starting time of drug administration. L , N – Q Representative images of IHC staining of the indicated xenograft tumors using the indicated antibodies ( L ). Magnified images of the boxed area are shown in the insets. Scale bars, 20 μm. Quantification of p-AKT1 (S473) intensity ( N ), p-UFL1 (T426) intensity ( O ), Ki-67-positive cell numbers ( P ), CC3-positive cell numbers ( Q ), in tumors. n = 5 mice per group. Data are presented as mean ± SD, and analyzed by one-way ANOVA with Tukey’s multiple comparisons test ( F , H , J , M – Q ) or two-way ANOVA with Tukey’s multiple comparisons test ( I ). Experiments were repeated three times ( A – H ) independently with similar results.

Article Snippet: For BODIPY staining, after treatment with 1 mM sodium citrate for 12 h in the absence or presence of 1 μM Fatostatin (MedChemExpress, HY-14452A), 45 μM C75 (MedChemExpress, HY-12364), 50 μM JR-AB2-011 (MedChemExpress, HY-14452A), 6 μM MK-2206 (MedChemExpress, HY-12464), or 20 μM DKM 2-93 (MedChemExpress, HY-122022), breast cancer cells were washed with PBS after fixation and stained with 0.25 μg/mL BODIPY 493/503 (Thermo Fisher, D3922) for 15 min.

Techniques: Phospho-proteomics, Staining, Immunohistochemistry