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: PF429242 , Selleckchem.com , Cat# S6418.

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: Journal of Translational Medicine

Article Title: Cathepsin H as a causal risk factor and therapeutic target in proliferative diabetic retinopathy

doi: 10.1186/s12967-025-07314-4

Figure Lengend Snippet: Docking results and interactions of cathepsin H with ( A ) SSR 69071 ( B ) PF429242 , in 3D and 2D formats

Article Snippet: Molecular dynamics simulations (100 ns) of the SSR 69071-CtsH and PF429242 -CtsH complexes confirmed stable dynamics: root mean square deviation (RMSD) plateaued below 0.2 nm after 40 ns (Fig. A), indicating conformational stability.

Techniques:

Journal: Journal of Translational Medicine

Article Title: Cathepsin H as a causal risk factor and therapeutic target in proliferative diabetic retinopathy

doi: 10.1186/s12967-025-07314-4

Figure Lengend Snippet: Effects of PF429242 and SSR 69071 on HUVEC healing rate, and lysosomal function. ( A ) Representative images of HUVECs at 0, 12, and 24 hours after treatment with different concentrations of PF429242 (veh, 2.5, 5, 10 μM); ( B ) Healing rate of HUVECs at 0 and 12 hours after treatment with different concentrations of PF429242 (veh, 2.5, 5, 10 μM); ( C ) Healing rate of HUVECs at 0 and 24 hours after treatment with different concentrations of PF429242 (veh, 2.5, 5, 10 μM); ( D ) Representative images of HUVECs at 0, 12, and 24 hours after treatment with different concentrations of SSR 69071 (veh, 0.01, 0.1, 1 μM); ( E ) Healing rate of HUVECs at 0 and 12 hours after treatment with different concentrations of SSR 69071 (0, 0.01, 0.1, 1 μM); ( F ) Healing rate of HUVECs at 0 and 24 hours after treatment with different concentrations of SSR 69071 (0, 0.01, 0.1, 1 μM); ( G ) Representative images of HUVECs treated with 10 μM PF429242 or 1 μM SSR 69071 for 4 hours, detected by LysoTracker Red staining (nuclei counterstained with Hoechst 33,342). ( H ) Representative images of HUVECs treated with 10 μM PF429242 or 1 μM SSR 69071 for 4 hours, captured by a Multi-SIM system, with lysosomes stained by LysoTracker Red and nuclei counterstained with Hoechst 33342. ( I ) Lysosomal area per cell of HUVECs treated with 10 μM PF429242 or 1 μM SSR 69071 for 4 hours. * p < 0.05; ** p < 0.01; *** p < 0.001

Article Snippet: Molecular dynamics simulations (100 ns) of the SSR 69071-CtsH and PF429242 -CtsH complexes confirmed stable dynamics: root mean square deviation (RMSD) plateaued below 0.2 nm after 40 ns (Fig. A), indicating conformational stability.

Techniques: Staining