jasplakinolide jas  (Santa Cruz Biotechnology)

Journal: Chinese Medical Journal

Article Title: Cell softness reveals tumorigenic potential via ITGB8/AKT/glycolysis signaling in a mice model of orthotopic bladder cancer

doi: 10.1097/CM9.0000000000002710

Figure Lengend Snippet: 3D Matrigel maintained the softness of tumor cells through F-actin/TRIM59. (A) Immunofluorescent staining of F-actin and TRIM59 in T24 cells cultured in flask, 3D matrix gels or 3D matrix gels containing Jas (20 mmol/L). The scale bar was 5 μm. (B) Western blotting of TRIM59 expression in MB49/T24 cells and TRIM59 KO MB49/T24 cells. (C) MB49/T24 cells and TRIM59 KO MB49/T24 cells were cultured in 3D matrix gel for 3 days. And these cells were digested and seeded in flask to check the colony formation in flask. (D) MB49/T24 cells and TRIM59 KO MB49/T24 cells were cultured in 3D matrix gel for 3 days. And these cells were digested and subcutaneously injected into NOD-SCID mice. After 20 days, the tumor formation was recorded ( n = 10). (E) T24 cells and TRIM59 KO T24 cells were cultured in 3D matrix gel for 3 days. And these cells were digested and examined by transwell for migration capacity. The scale bar was 50 μm. (F) The overall survival of bladder cancer patients with high/low TRIM59 expression from TCGA ( n = 128 in each group, HR (95% CI) = 0.7402 (0.5514–0.9944), P = 0.044). Values were shown as mean ± SD. * P <0.01. DAPI: 4′,6-diamidino-2-phenylindole; KO: Knock out; VEC: Vehicle; NOD-SCID: Non-obese diabeticsevere combined immune deficiency; TCGA: The Cancer Genome Atlas; HR: Hazard ratio; CI: Confidence interval; Jas: Jasplakinolide; SD: Standard deviation.

Article Snippet: Jasplakinolide (Jas) and AKT inhibitor (3-chloroacetyl-indole) 3CAI were purchased from MedChemExpress (South Brunswick Township, NJ, USA).

Techniques: Staining, Cell Culture, Western Blot, Expressing, Injection, Migration, Knock-Out, Standard Deviation

Journal: Communications Biology

Article Title: Temporal regulation of BMP2 growth factor signaling in response to mechanical loading is linked to cytoskeletal and focal adhesion remodeling

doi: 10.1038/s42003-024-06753-x

Figure Lengend Snippet: a Representative images showing the dynamic protrusion remodeling of LifeAct-GFP expressing hFOBs during a period of three minutes before and after Jas treatment. Remodeling dynamics are visualized by overlaying all cell outlines stained from blue to pink according to the frame number. The overlay was prepared using the QuimP ImageJ plugin. The kymographs taken along the red lines illustrate the different protrusion dynamics. Quantification of the mean change in protrusion area per time frame before and after Jas treatment (20–45 min after). Change in protrusion area after Jas treatment was normalized to the untreated control (=before treatment) ( n = 12). b Human FOBs seeded into collagen scaffolds were incubated for 3 h in a starvation medium supplemented with 0.1 µM Jasplakinolide (Jas) or the same amount of DMSO. Subsequently, scaffolds were subjected to BMP2 stimulation (B, 5 nM), mechanical loading (L, 1 Hz, 10%), or a combination of both (B/L) for 90 min. Cells were fixated and stained for phospho-Paxillin (green), F-actin with phalloidin (red), and nuclei with DAPI (blue). Representative confocal images of stained hFOBs are depicted. Scale bar represents 50 µm. c Total number of phospho-Paxillin positive FA per cell and d percentage of cells with medium and large FAs (medium = 0.7–1 µm 2 , large >1 µm 2 ) were evaluated in ImageJ (in total >100 cells per condition, n = 4). For the analysis of e Smad1/5/8 phosphorylation by western blot, or f ID1 expression by qPCR, cells were lysed in the respective assay buffer after the bioreactor experiment ( n ≥ 3 of ≥3 independent experiments). Statistical significance was calculated using a two-sided Mann–Whitney U test ( # p < 0.1, * p < 0.05, **p < 0.01, *** p < 0.001).

Article Snippet: To investigate whether load-induced actin cytoskeleton rearrangement processes are important for the integration of mechanical signals into the BMP pathway, the actin-binding macrocyclic peptide Jasplakinolide (Jas, J4580, Sigma-Aldrich), an inhibitor of actin depolymerization , was used as an agent to stabilize actin filaments.

Techniques: Expressing, Staining, Control, Incubation, Western Blot, MANN-WHITNEY