fam caspase activity kit  (Novus Biologicals)

Journal: Oncology Reports

Article Title: PINK1-mediated mitophagy enhances breast cancer proliferation through metabolic reprogramming

doi: 10.3892/or.2026.9117

Figure Lengend Snippet: Analysis of glycolysis-associated metabolite level changes in MCF-7 and MDA-MB-231 breast cancer cells. Levels of (A) PGK2, (B) pyruvate and (C) acetyl-CoA in MCF-7 cells, and (D) PGK2, (E) pyruvate and (F) acetyl-CoA in MDA-MB-231 cells transfected with vector control, PINK1-OE or PINK1-OE + siPGK2. Quantitative analysis was performed using ELISA in cells under the three indicated conditions. Statistical analysis was performed using a one-way ANOVA followed by Tukey's post-hoc test. Data are represented as the mean ± SEM from at least three independent experiments. ***P<0.001. PGK2, phosphoglycerate kinase 2; PINK1, PTEN-induced kinase 1; OE, overexpression; si, small interfering.

Article Snippet: Acetyl-CoA content was assessed using the Acetyl-CoA ELISA Kit (Wuhan Elabscience Biotechnology Co., Ltd.; cat. no. E-BC-F046-48T-ELS).

Techniques: Transfection, Plasmid Preparation, Control, Enzyme-linked Immunosorbent Assay, Over Expression

Journal: The Journal of Biological Chemistry

Article Title: Siah2 regulates lipid uptake in adipose tissue macrophages

doi: 10.1016/j.jbc.2026.111380

Figure Lengend Snippet: Lysosomal program is upregulated in ATMs independent of Siah2 . A , relative mRNA expression of lysosomal genes in fl/fl and Siah2 MacKO BMDMs and BM-ATMs. B , representative Western blot of lysosomal enzymes relative to β-actin in fl/fl and Siah2 MacKO BMDMs and BM-ATMs. The β-actin panel is the same as used in F . The blot shown in F was reprobed for the CTSK image in B . C , enzyme activity of CTSK and LAL in fl/fl and Siah2 MacKO BMDMs and BM-ATMs. D and E , relative mRNA expression of Atgl ( D ) and Hilpda ( E ) in fl/fl and Siah2 MacKO BMDMs and BM-ATMs. F , representative Western blot of ATGL and HILPDA relative to β-actin in fl/fl and Siah2 MacKO BMDMs and BM-ATMs. G , basal glycerol release in fl/fl and Siah2 MacKO BMDMs and BM-ATMs treated with vehicle or 50 μM Atglistatin or 0.1 μM Lalistat-1 for 24 h. H , glycerol release in fl/fl and Siah2 MacKO BM-ATMs treated with vehicle or 50 μM Atglistatin or 0.1 μM Lalistat-1 for 24 h with or without 0.5 mM IBMX (activated lipolysis). I and J , representative confocal images ( I ) and quantification ( J ) of fl/fl and Siah2 MacKO BM-ATMs stained for BODIPY 493/503 (LD, green ), LysoTracker (lysosomes, magenta ), and DAPI (nuclei, blue ) showing colocalization of LD and lysosomes. The scale bar represents 10 μm. Red signals of LysoTracker were converted to magenta using the LUT function in ImageJ. Statistics are reported as mean ± SD using an unpaired t test with Welch’s correction, and each dot denotes technical replicates representative of three to five independent experiments. A , D , E , G , H , and J , statistics are reported as mean ± SD using two-way ANOVA with Tukey’s multiple comparisons test ( C ). p Values are indicated on the graphs. ATGL, adipose triglyceride lipase; ATM, adipose tissue macrophage; BM-ATM, bone marrow–derived ATM; BMDM, bone marrow–derived macrophage; CTSK, cathepsin K; DAPI, 4′,6-diamidino-2-phenylindole; fl/fl, Siah2 flox/flox ; HILPDA, hypoxia-inducible LD-associated protein; IBMX, 3-isobutyl-1-methylxanthine; LAL, lysosomal acid lipase; LD, lipid droplet; MacKO, Siah2 MacKO ; Siah2, seven in absentia homolog 2.

Article Snippet: CTSK activity in BMDMs and BM-ATMs was measured using the Cathepsin K assay kit (Novus Biologicals, catalog no.: NBP2-54842) according to the manufacturer’s protocol.

Techniques: Expressing, Western Blot, Activity Assay, Staining, Derivative Assay