anti β actin mouse monoclonal antibody  (Cell Signaling Technology Inc)

Journal: Cancer Pathogenesis and Therapy

Article Title: Metabolic pathways and chemotherapy resistance in acute myeloid leukemia (AML): Insights into Enoyl-CoA hydratase domain-containing protein 3 ( ECHDC3 ) as a potential therapeutic target

doi: 10.1016/j.cpt.2025.08.002

Figure Lengend Snippet: Changes in mitochondrial function following ECHDC3 knockdown. (A) TMRE staining results based on ECHDC3 -knockdown cells. siNC cells emitted bright red-orange fluorescence. Cells treated with a mitochondrial membrane-potential disrupter, CCCP, showed very weak or complete absence of red-orange fluorescence. The average fluorescence intensity of the cells was calculated and quantitatively analyzed. (B–C) mtDNA copy number ( MT–CO1 and MT–CO2 ) was quantified via quantitative RT-PCR; (D) Quantitation of mitochondrial SOD activity, wherein SOD activity decreased in ECHDC3 -knockdown cells. (E) Mitophagy biomarkers were detected via western blotting. β-Actin was used as a control. (F–I) Quantitation of the mitophagy pathway protein. Values were presented as mean ± standard error. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001. CCCP: Carbonyl cyanide m-chlorophenyl hydrazone; ECHDC3 : Enoyl-CoA hydratase domain-containing protein 3; mtDNA: Mitochondrial DNA; RT-PCR: Real-time polymerase chain reaction; SOD: Superoxide dismutase; TMRE: Tetramethyl rhodamine ethyl ester.

Article Snippet: Western blotting was performed to determine the expression of mitochondrial proteins, using the Mitophagy Antibody Sampler Kit (Cat# 43110, Cell Signaling Technology [CST], MA, USA) and an anti-β-actin mouse monoclonal antibody (Cat# 3700, CST, MA, USA).

Techniques: Knockdown, Staining, Fluorescence, Membrane, Quantitative RT-PCR, Quantitation Assay, Activity Assay, Western Blot, Control, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction

Journal: Tumour Virus Research

Article Title: HPV18E6 and CDK5 virus-host interaction is a prospective therapeutic target for HPV-positive cervical cancer

doi: 10.1016/j.tvr.2026.200339

Figure Lengend Snippet: CP681301 inhibits CDK5 and 18E6 in HeLa cell line, and disrupted the CDK5-HPV18E6 association directly. (A) (i). CP681301 disrupted the CDK5-18E6 protein complex. The inhibition of CP681301 was detected using the GST-pull down assay, where CP681301 was incubated with the purified GST-18E6 protein and CDK5 protein for 2h. After extensive washing, the bound CDK5 protein was detected via Western blotting using an anti-CDK5 antibody. The immunoblot (IB) on the upper panel shows the interaction of CDK5 with GST-18E6, while the lower panel shows the Ponceau S stained of the blot. (ii). The bar graph shows the quantification of the relative level of CDK5 to GST-18E6 protein indicated from 3 independent experiments (n = 3). Quantitation was performed using ImageJ software, and the statistical analysis was performed using Prism. (B). The summarisation of the CP681301 binding affinity value with 16E6, 18E6 and CDK5. (C) (i). The immunoblot shows the protein levels of HPV18E6 and CDK5 at different concentrations of CP681301. HeLa cells were treated with different concentrations of CP681301 (0.2 to 3.2 μM) or DMSO as indicated. Total cell lysates were subjected to western blotting with anti-CDK5, anti-18E6, and anti-β-actin antibodies. (ii). The EC50 of 18E6 in the treatment of CP681301. (iii). The EC 50 of CDK5 in the treatment of CP681301. The percentage of CDK5 and 18E6 protein levels was analyzed using the nonl i near regression function in GraphPad Prism 8.

Article Snippet: The transferred membranes were incubated overnight at 4 °C with the following primary antibodies: monoclonal rabbit anti-HA (Cell Signaling Technology), monoclonal mouse anti-p53 (Santa Cruz), monoclonal mouse anti-HPV18E6 (Santa Cruz), monoclonal mouse anti-CDK5 (Santa Cruz), and monoclonal mouse anti-β-actin (Santa Cruz).

Techniques: Inhibition, Pull Down Assay, Incubation, Purification, Western Blot, Staining, Quantitation Assay, Software, Binding Assay

Journal: Neoplasia (New York, N.Y.)

Article Title: BPGM as an intrinsic brake to constrain metastasis through phospho-epigenetic-mediated carnitine biosynthesis suppression

doi: 10.1016/j.neo.2026.101299

Figure Lengend Snippet: Overexpression of BPGM inhibits tumor metastasis in vitro and in vivo . (A) Silencing BPGM promoted migration of tumor cells. Tumor cells stably expressing shBPGM and its control cells (shCtrl) were examined. Scale bar, 200 μm. (B) Overexpressing BPGM suppressed migration of tumor cells. Tumor cells stably expressing BPGM and its control cells (Ctrl) were examined. Scale bar, 200 μm. (C) The mRNA levels of MMP2 and MMP9 reduced in BPGM-overexpressing tumor cells. SK-HEP-1 cells stably expressing BPGM (BPGM-OE) and its ctrl cells were employed to detect the mRNA levels of MMP2 and MMP9 by qPCR analysis. β-actin was used as an internal control. (D-E) Xenografts of stable Bpgm-overexpressing cells displayed a lower rate of liver and lung metastasis and less metastatic nodules in the liver. For (D), Hepa-Ctrl ( n = 7) and Hepa-BPGM sublines ( n = 6) were inoculated under the capsule of the left hepatic lobe of C57BL/6 mice. Upper panel, a schematic diagram of orthotopic hepatic implantation model. The number of metastatic rate and nodules is shown (D, lower panel). Scale bar, 1 cm. Hematoxylin-eosin staining was performed on serial sections of livers (E, left panel) and lungs (E, right panel) to detect the metastatic nodules. The red arrows indicated the metastatic nodules. Scale bar in left panel, 200 µm; Scale bar in right panel, 100 μm. (F-H) Overexpressing of BPGM inhibited lung metastasis of tumor xenografts. Scale bar in F, 1 cm. B16-F10 cells transfected with Ctrl ( n = 6) or BPGM-OE ( n = 6) was injected into the tail vein of C57BL/6 mice, respectively. Upper panel in F, a schematic diagram of lung metastasis model by tail vein injection. H&E staining of lung sections was performed to observe metastatic foci (G). For G, scale bar in left panel, 500 μm; scale bar in right panel, 200 μm. The number of melanoma nodules and lung metastasis is shown in H. (I) The model deciphers the inhibitory role of BPGM in tumor metastasis. Error bar: mean ± SEM. P -values are labeled above the bar chart.

Article Snippet: The antibodies used included mouse antibody against β-actin (BM0627, Boster, Wuhan, China), rabbit antibody against BPGM (17173-1-AP, Proteintech), EZH2 (F0281, Selleck), phospho-EZH2 (Thr345) (TA3584S, Abmart, Shanghai, China), phospho-CDK1 (Thr14) (AP1465, Abclonal, Wuhan, China), ubiquitin (10201-2-AP, Proteintech), HIF1α (36169, Cell Signaling Technology, CST, Beverly, MA, USA), H3K4me3 (91264, Active Motif), H3K79me3 (cat 49-1020, Thermos Fisher), H3K9me3 (61014, Active Motif), H3K27me3 (91168, Active Motif) and Histone 3 (F0057, Selleck).

Techniques: Over Expression, In Vitro, In Vivo, Migration, Stable Transfection, Expressing, Control, Staining, Transfection, Injection, Labeling

Journal: iScience

Article Title: Single-cell transcriptomic analyses reveal angiogenic vascular responses to chronic cerebral hypoperfusion

doi: 10.1016/j.isci.2026.115331

Figure Lengend Snippet: Tip cells show enhanced function in angiogenesis with an upregulation in Apln after VD (A) Volcano plot showing a total of 1,472 up- (right) and downregulated (left) DEGs (|log2 FC| > 1 and Bonferroni-adjusted p value <0.05) of tip cells versus other EC subclusters in ACAS brains. (B) Gene Ontology (GO) enrichment analysis of DEGs identified in (A). Five major classes of GO terms were identified. Top genes of each plotted GO term are shown in the right. Dot colors representing the significance level (−log10(adjusted p value)) and dot sizes indicating the number of genes in each category. (C) UMAP plot showing expression of Apln and Aplnr in ACAS and sham cells. (D) UMAP plots depicting the co-expression visualization of Apln and Aplnr in ECs. The colors represent scaled expression levels. (E) Representative immunostaining images of Apln (red), tip cell marker CD93 (green), endothelial cell marker CD31 (magenta), and DAPI nuclear staining (blue) in the brain 35 days after ACAS. The Apelin + CD93 + CD31 + tip cells in white squares are enlarged and 3D reconstructed to show colocalization. (F) Representative images of western blots for Apln in brain 35 days after ACAS or sham operation. (G) Quantification of protein levels of Apln. β-actin was used as a loading control. n = 4–7/group. Student’s t test. Data are shown as mean ± SEM. ∗ p < 0.05. Scale bars are indicated in images.

Article Snippet: The primary antibodies used include Apln (1:500, Cat# PA5-114860; Invitrogen, Carlsbad, CA) and β-actin (1:5000, Cat# MAB8929; R&D systems, Minneapolis, MN).

Techniques: Expressing, Immunostaining, Marker, Staining, Western Blot, Control