Journal: Cells

Article Title: Corosolic Acid Attenuates the Invasiveness of Glioblastoma Cells by Promoting CHIP-Mediated AXL Degradation and Inhibiting GAS6/AXL/JAK Axis

doi: 10.3390/cells10112919

Figure Lengend Snippet: Involvement of GAS6-associated cascade in CA-inhibited migration and invasion of GBM cells. ( A ) Cells were treated with CA (20 μM) and then lysed for immunodetection of the indicated targets by Western blotting. ( B , C ) Cells were treated with CA (20 μM), GAS6 (100 ng/mL), or a combination of CA and GAS6, and then lysed for immunodetection of the indicated targets ( B ) or F-actin ( C ) by Western blotting. ( D ) Cells were treated with CA (20 μM), GAS6 (100 ng/mL) or a combination of CA and GAS6, then subjected to migration and invasion assay. Chemiluminescence signal was semi-quantitated by densitometric analysis, and GAPDH was used as an internal control. * and ** p < 0.05 and 0.01, respectively, compared with the control (DMSO-treated cells). # p < 0.05 compared with CA alone. Images were acquired using a light microscope at 200× magnification.

Article Snippet: Recombinant Human GAS6 Protein (Rh-GAS6) was purchased from R&D Systems, Inc (Minneapolis, MN, USA)

Techniques: Migration, Immunodetection, Western Blot, Invasion Assay, Control, Light Microscopy

Journal: Cells

Article Title: Corosolic Acid Attenuates the Invasiveness of Glioblastoma Cells by Promoting CHIP-Mediated AXL Degradation and Inhibiting GAS6/AXL/JAK Axis

doi: 10.3390/cells10112919

Figure Lengend Snippet: Molecular docking analysis and proposed mechanism for CA-inhibited invasiveness of GBM cells. ( A ) Superposition of GAS6 (green) and template CA (cyan), hydro-gen bonding interactions with GAS6 and CA. Binding affinity: −7.6 kcal/mol. ( B ) Superposition of AXL (green) and template CA (cyan), hydrogen bonding interactions with AXL and CA. Binding affinity: −5.6 kcal/mol. Interacting amino acid residues: Asn677, Arg676, Leu542 and Asp672. ( C ) Proposed mechanism for CA-inhibited invasiveness of GBM cells.

Article Snippet: Recombinant Human GAS6 Protein (Rh-GAS6) was purchased from R&D Systems, Inc (Minneapolis, MN, USA)

Techniques: Binding Assay

Journal: Molecular Cancer Therapeutics

Article Title: Evaluation of the Role of AXL in Fusion-positive Pediatric Rhabdomyosarcoma Identifies the Small-molecule Inhibitor Bemcentinib (BGB324) as Potent Chemosensitizer

doi: 10.1158/1535-7163.mct-23-0285

Figure Lengend Snippet: Figure 2. Gas6 stimulates Axl signaling in FP-RMS cells. A, Representative Western blot images showing AXL expression in IC-pPDX-104 and Rh41 WT, KO (sgAXL-1) and OE lines. GAPDH was used as loading control. B, Morphologic appearance of IC-pPDX-104 and Rh41 cells with AXL WT, KO and OE. Scale bar, 70 mm. C, Schematic representation of the AXL signaling pathway. D, Representative Western blot images of AXL WT, KO (sgAXL-1) and OE cells stimulated with 400 ng/mL GAS6 for 10 minutes. E, Quantification of Western blots measuring the phosphorylation of downstream targets of AXL in AXL WT, KO (sgAXL-1) and OE cells stimulated with 400 ng/mL GAS6 for 10 minutes. Data are represented as mean SEM of the indicated number of independent biological replicates; ordinary two-way ANOVA with Sidak multiple comparisons test. ns, nonsignificant.

Article Snippet: The next day, we stimulated cells with the indicated concentration of GAS6 (R&D Systems, 885-GSB-050) for 10 minutes at 37 C, and processed them for Western blot analysis.

Techniques: Western Blot, Expressing, Control, Phospho-proteomics

Journal: Molecular Cancer Therapeutics

Article Title: Evaluation of the Role of AXL in Fusion-positive Pediatric Rhabdomyosarcoma Identifies the Small-molecule Inhibitor Bemcentinib (BGB324) as Potent Chemosensitizer

doi: 10.1158/1535-7163.mct-23-0285

Figure Lengend Snippet: Figure 5. Axl contributes to the migratory phenotype of Rh41 cells. A–C, Effect of AXL expression levels on the migratory phenotype of Rh41 cells. Representative images of Rh41 AXL WT (sgAAVS1), KO (sgAXL-1, sgAXL-2) and OE cells (A) and quantification (B) of wound-healing assay. Scale bar, 100 mm. C, Wound area remaining after 20 hours. D–F, Effect of bemcentinib on the migratory phenotype of Rh41 cells. Representative images (D) and quantification (E) of wound-healing assay performed on Rh41 WT cells treated with the indicated concentrations of bemcentinib. Scale bar, 100 mm. F, Wound area remaining after 12 hours in Rh41 cells. G and H, Effect of bemcentinib and AXL stimulation with GAS6 on the migratory phenotype of AXL KO cells. G, Quantification of wound-healing assay performed on Rh41 AXL WT (sgAAVS1) and KO (sgAXL-1, sgAXL-2) cells treated with the indicated concentrations of bemcentinib or with 400 ng/mL GAS6. H, Wound area remaining after 15 hours in Rh41 cells. Data are represented as mean þ SEM of n ¼ 3 biological replicates. Ordinary one-wayANOVA with Dunnett multiple comparison test againstthe control group (WT or 0 mmol/L bemcentinib).

Article Snippet: The next day, we stimulated cells with the indicated concentration of GAS6 (R&D Systems, 885-GSB-050) for 10 minutes at 37 C, and processed them for Western blot analysis.

Techniques: Expressing, Wound Healing Assay, Comparison, Control