Journal: iScience

Article Title: Targeting host lipogenesis with a diarylamide inhibitor disrupts SARS-CoV-2 replication

doi: 10.1016/j.isci.2026.117111

Figure Lengend Snippet: NCOA1 is a host factor targeted by compound 10 to regulate lipid metabolic programs (A) Validation of DARTS assay by western blot in Huh-7 cell lysates. Compd. 10 (50 μM) treatment increases the protease resistance of NCOA1 protein. (B and C) CETSA analysis reveals that compd. 10 (50 μM) induced thermal stabilization of NCOA1 is abolished by D276A and R281A mutations. Experiments were performed in HEK293T cells. (D and E) Western blot analysis shows a dose-dependent reduction of NCOA1 protein levels by compd. 10 . Data were analyzed by one-way ANOVA followed by Dunnett’s test (∗∗ p < 0.01). (F and G) CHX chase assay demonstrates accelerated degradation of NCOA1 in the presence of compd. 10 (8 μM). Student’s t test was used for statistical analysis (∗ p < 0.05, ∗∗ p < 0.01). (H and I) CHX chase analysis shows reduced stability of PPARγ protein upon compd. 10 (8 μM) treatment. Student’s t test was used for statistical analysis (∗∗ p < 0.01). (J and K) Western blot analysis shows that autophagy-lysosome inhibitors (3-MA, BafA1), but not the proteasome inhibitor MG132, rescue NCOA1 protein levels reduced by compd. 10 (8 μM), indicating lysosome-dependent degradation. Data were analyzed by two-way ANOVA followed by Šídák’s test (∗∗ p < 0.01, ∗∗∗ p < 0.001). Data are presented as mean ± SD from at least three independent experiments.

Article Snippet: 3-methyladenine (3-MA) , MCE , Cat# HY-19312.

Techniques: Biomarker Discovery, Western Blot

Journal: iScience

Article Title: BUB1 regulates cell proliferation and apoptosis in cervical cancer by AKT signaling and autophagosome-lysosome fusion via ACTN1

doi: 10.1016/j.isci.2026.117040

Figure Lengend Snippet: BUB1 KD suppressed autophagosome-lysosome fusion, induced ROS production, and then increased cell apoptosis of CC cells (A) Expression of autophagy-related proteins in BUB1 KD or control HeLa and Ca Ski cells were determined by western blot assay. β-Actin was used as a loading control. (B) Quantitation of western blot assay bands shown in (A) using ImageJ. Statistical analysis was performed using Student’s t test. (C) Representative image of Ca Ski cells after BUB1 KD stained with LC3B. Scale bars, 100 μm. (D) Confocal fluorescence imaging was used to determine autophagosome maturation of Ca Ski cell transfected with sh-NC and sh-BUB1. Scale bars, 50 μm. The colocalization analysis was expressed as Pearson’s correlation coefficient, measured at least 60 cells of three independent experiments. (E) Representative image of Ca Ski cells after BUB1 KD stained with LysoTracker red showing the number of acidic lysosomes. Scale bars, 50 μm. (F) Confocal fluorescence imaging was used to determine autophagosome-lysosome encounter of Ca Ski cells transfected with sh-NC and sh-BUB1. Scale bars, 50 μm. The colocalization analysis was expressed as Pearson’s correlation coefficient, measured at least 60 cells. (G) Expression of SQSTM1, LC3-I, and LC3-II in Ca Ski cells treated with the inhibitors Baf-A1, CQ, and 3-MA were determined by western blot assay. β-Actin was used as a loading control. (H) Flow cytometry assay was used to detect cell apoptosis in Ca Ski cells treated with the inhibitors Baf-A1, CQ, and 3-MA. (I) Flow cytometry assay was used to detect ROS production in Ca Ski cells treated with the inhibitors Baf-A1, CQ, and 3-MA. (J) Flow cytometry assay was used to detect cell apoptosis in Ca Ski cells treated with NAC, the inhibitor of ROS. All experiments were repeated three times at least. The data were presented as mean ± SEM, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.005, and ∗∗∗∗ p < 0.001. Baf-A1, bafilomycin-A1; CQ, chloroquine; 3-MA, 3-methyladenine; ROS, reactive oxygen species; NAC, N-acetyl-L-cysteine.

Article Snippet: 2OH-BNPP1 (HY-102081), Bafilomycin A1 (HY-100558), Chloroquine (HY-17589A), 3-Methyladenine (HY-199312) was obtained from MedChemExpress (NJ, USA).

Techniques: Expressing, Control, Western Blot, Quantitation Assay, Staining, Fluorescence, Imaging, Transfection, Flow Cytometry