hspa8 protein  (MedChemExpress)

Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

Article Title: Pristimerin Promotes Ubiquitination of HSPA8 and Activates the VAV1/ERK Pathway to Suppress TNBC Proliferation.

doi: 10.1002/advs.202413174

Figure Lengend Snippet: Figure 4. Pristimerin directly targets HSPA8. (A) Schematic depicting the rough flow of the DARTS-MS assay. (B) Western blot of CETSA for HSPA8 with pristimerin or DMSO treatment for 1 h in TNBC cells. (C) SDS-PEAG of the HSPA8 recombinant protein. (D) The binding affinity of HSPA8 recombinant protein with pristimerin was detected by SPR analysis. (E) The binding of HSPA8 with pristimerin was simulated by molecular docking. (F–H) The expression of HSPA8 in normal and tumor tissues were analyzed through UALCAN website (https://ualcan.path.uab.edu/index.html) and BCa Gene- Expression Miner website (http://bcgenex.ico.unicancer.fr/BC-GEM/GEM-Accueil.php?js = 1). (I) The prognostic significance of HSPA8 in BCa (TCGA

Article Snippet: Surface Plasmon Resonance (SPR): The PlexArray HT A100 system was employed to measure the binding affinity between HSPA8 recombinant protein (MedChemExpress) and pristimerin/doxorubicin/ docetaxel.

Techniques: Western Blot, Recombinant, Binding Assay, Expressing, Gene Expression

Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

Article Title: Pristimerin Promotes Ubiquitination of HSPA8 and Activates the VAV1/ERK Pathway to Suppress TNBC Proliferation.

doi: 10.1002/advs.202413174

Figure Lengend Snippet: Figure 5. The anticancer activity of pristimerin relies on HSPA8. (A) The level of HSPA8 was determined in HSPA8 knockdown cells combined with pristimerin treatment. (B-C) HSPA8 knockdown cells were treated with DMSO or pristimerin for 48 h, and then the cell viability and colony formation ability were assessed through CCK-8 (B) and colony formation assays (C), respectively. (D) HSPA8 knockdown cells were treated with DMSO or pris- timerin for 48 h, and then the autophagosomes and autolysosomes were examined by immunofluorescence. (E) HSPA8 knockout cells (4T1-shNC and 4T1-shHSPA8) were injected subcutaneously into the left breast fat pad of BALB/c mice and palpable tumors were allowed to develop for 7 days. Mice were randomly divided into four groups (n = 6) and treated with vehicle control (0.5% CMC-Na) or 1 mg kg−1 pristimerin every other day. The tumor size was measured at indicated time intervals and tumors were imaged at the end of treatment. (F) The expressions of HSAP8, VAV1 and p-ERK were detected by immunohistochemical assay. Scale bar = 50 μm Bars, SDs; *0.01 < p < 0.05, **0.001 < p < 0.01, and ***p < 0.001.

Article Snippet: Surface Plasmon Resonance (SPR): The PlexArray HT A100 system was employed to measure the binding affinity between HSPA8 recombinant protein (MedChemExpress) and pristimerin/doxorubicin/ docetaxel.

Techniques: Activity Assay, Knockdown, CCK-8 Assay, Knock-Out, Injection, Control, Immunohistochemical staining

Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

Article Title: Pristimerin Promotes Ubiquitination of HSPA8 and Activates the VAV1/ERK Pathway to Suppress TNBC Proliferation.

doi: 10.1002/advs.202413174

Figure Lengend Snippet: Figure 6. Pristimerin attenuates HSPA8-mediated VAV1 degradation. (A,B) After treatment with different concentrations of pristimerin for 48 h, the HSPA8 expression was tested by Western blot (A) and qPCR (B). (C) Western blot was used to analyze HSPA8 expression in TNBC cells exposed to 100 μg mL−1 CHX in the presence or absence of pristimerin at the indicated times. (D) TNBC cells were transfected with Ub-HA plasmid for 36 h, and then treated with 1 μm pristimerin and 5 μm MG-132 for 24 h, followed by immunoprecipitation with anti-HSPA8 antibody, and finally, HA levels were detected by Western blot. (E) HSPA8 overexpression cells were treated with pristimerin or DMSO for 48 h, cell lysis was collected and then immunoprecipitation

Article Snippet: Surface Plasmon Resonance (SPR): The PlexArray HT A100 system was employed to measure the binding affinity between HSPA8 recombinant protein (MedChemExpress) and pristimerin/doxorubicin/ docetaxel.

Techniques: Expressing, Western Blot, Transfection, Plasmid Preparation, Immunoprecipitation, Over Expression, Lysis

Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

Article Title: Pristimerin Promotes Ubiquitination of HSPA8 and Activates the VAV1/ERK Pathway to Suppress TNBC Proliferation.

doi: 10.1002/advs.202413174

Figure Lengend Snippet: Figure 9. Schematic representation of the mechanisms by which pristimerin inhibits TNBC progression. In TNBC cells, HSPA8 facilitates the degradation of VAV1, thereby inhibiting the activation of the ERK pathway, suppressing cellular autophagy, and regulating the balance between cell proliferation and apoptosis. However, in the presence of pristimerin, it directly targets HSPA8, promoting its ubiquitination and degradation. This further inhibits the degradation of the downstream client protein VAV1, leading to the activation of the ERK pathway and mediating cellular autophagy and apoptosis. Ultimately, this leads to the suppression of cell proliferation, cell migration, and drug resistance.

Article Snippet: Surface Plasmon Resonance (SPR): The PlexArray HT A100 system was employed to measure the binding affinity between HSPA8 recombinant protein (MedChemExpress) and pristimerin/doxorubicin/ docetaxel.

Techniques: Activation Assay, Ubiquitin Proteomics, Migration