Journal: International Journal of Biological Sciences

Article Title: Targeted Induction of Cancer Cell Necroptosis Potentiates Anti-PD-1 Immunotherapy via CD80 Activation

doi: 10.7150/ijbs.121690

Figure Lengend Snippet: CL-387785 Inhibits the Proliferation of Melanoma and Lung Cancer Cells. (A) Chemical structure of the small-molecule inhibitor CL-387785. (B) CCK-8 assays were conducted to determine the IC 50 of CL-387785 and its effects on SK-MEL-5, A375 (melanoma), A549, and H1299 (lung cancer) cells. n=4-6. (C) Colony formation assays were performed on melanoma and lung cancer cell lines to evaluate the long-term anti-proliferative effects of CL-387785. n=3. (D) Quantitative analysis of the colony formation assays. (E) The effect of CL-387785 on cell cycle distribution was evaluated via flow cytometry assay. n=3.

Article Snippet: SK-MEL-5, SK-MEL-28, A375, B16-F10, A549, H1299 and PIG1 cells (3×10 3 cells per well) were inoculated into a 96-well plate, cultured with complete medium supplemented with different concentrations of CL-387785 (HY-10325; MedChemExpress, USA), and then incubated at 37 °C in a thermostatic incubator containing 5% CO 2 for 24, 48, or 72 hours.

Techniques: CCK-8 Assay, Flow Cytometry

Journal: International Journal of Biological Sciences

Article Title: Targeted Induction of Cancer Cell Necroptosis Potentiates Anti-PD-1 Immunotherapy via CD80 Activation

doi: 10.7150/ijbs.121690

Figure Lengend Snippet: CL-387785 Inhibits Migration and Invasion in Melanoma and Lung Cancer Cells. (A) Wound healing assays were performed to assess the effect of CL-387785 on the migration of SK-MEL-5, A375 (melanoma), A549, and H1299 (lung cancer) cells. ×20 magnification. n=3. (B) Quantitative analysis of the wound healing assays. (C) Transwell assays were conducted to evaluate the inhibitory effect of CL-387785 on invasion. ×20 magnification. n=3. (D) Quantitative analysis of the cell invasion results.

Article Snippet: SK-MEL-5, SK-MEL-28, A375, B16-F10, A549, H1299 and PIG1 cells (3×10 3 cells per well) were inoculated into a 96-well plate, cultured with complete medium supplemented with different concentrations of CL-387785 (HY-10325; MedChemExpress, USA), and then incubated at 37 °C in a thermostatic incubator containing 5% CO 2 for 24, 48, or 72 hours.

Techniques: Migration

Journal: International Journal of Biological Sciences

Article Title: Targeted Induction of Cancer Cell Necroptosis Potentiates Anti-PD-1 Immunotherapy via CD80 Activation

doi: 10.7150/ijbs.121690

Figure Lengend Snippet: CL-387785 Induces Necroptosis in Tumor Cells. (A) ROS levels in SK-MEL-5, A375 (melanoma), A549, and H1299 (lung cancer) cells were detected after treatment with CL-387785. n=3. (B) The morphological characteristics of SK-MEL-5, A375, A549, and H1299 cells treated with CL-387785 were observed by light microscopy. ×20 magnification. (C) Necroptotic features of melanoma cells were further visualized using transmission electron microscopy after CL-387785 treatment. (D) Hoechst/PI staining was performed to assess necroptotic cell death in melanoma and lung cancer cells treated with CL-387785. Quantitative analysis of the Hoechst/PI staining is shown in the image. ×20 magnification. n=3. (E) Western blot analysis was used to assess the protein levels of phosphorylated RIPK1 (p-RIPK1), RIPK1, phosphorylated MLKL (p-MLKL), MLKL, and phosphorylated JNK (p-JNK) in SK-MEL-5, A375, A549, and H1299 cell lysates. (F) Rescue effects of NEC-1 and NSA on CL-387785-induced cell death in A375 and A549 cells. n=4.

Article Snippet: SK-MEL-5, SK-MEL-28, A375, B16-F10, A549, H1299 and PIG1 cells (3×10 3 cells per well) were inoculated into a 96-well plate, cultured with complete medium supplemented with different concentrations of CL-387785 (HY-10325; MedChemExpress, USA), and then incubated at 37 °C in a thermostatic incubator containing 5% CO 2 for 24, 48, or 72 hours.

Techniques: Light Microscopy, Transmission Assay, Electron Microscopy, Staining, Western Blot

Journal: PLoS ONE

Article Title: Intracellular galectin-7 expression in cancer cells results from an autocrine transcriptional mechanism and endocytosis of extracellular galectin-7

doi: 10.1371/journal.pone.0187194

Figure Lengend Snippet: Expression of galectin-7 was measured by Western blot at different times following stimulation with rhGal-7.

Article Snippet: Anti-human galectin-7 was purchased from R&D Systems (Minneapolis, MN, USA) while anti-lamin A/C, anti-Cox IV and anti-β-tubulin were obtained from Cell Signaling (Danvers, MA, USA).

Techniques: Expressing, Western Blot

Journal: PLoS ONE

Article Title: Intracellular galectin-7 expression in cancer cells results from an autocrine transcriptional mechanism and endocytosis of extracellular galectin-7

doi: 10.1371/journal.pone.0187194

Figure Lengend Snippet: Western blot analysis in MDA-MB-231 cells showing expression of cytosolic and mitochondrial galectin-7 15 min after stimulation with rhGal-7 at 0°C or 37°C. Tubulin and CoxIV were used as controls for cytosolic and mitochondrial extracts respectively.

Article Snippet: Anti-human galectin-7 was purchased from R&D Systems (Minneapolis, MN, USA) while anti-lamin A/C, anti-Cox IV and anti-β-tubulin were obtained from Cell Signaling (Danvers, MA, USA).

Techniques: Western Blot, Expressing

Journal: PLoS ONE

Article Title: Intracellular galectin-7 expression in cancer cells results from an autocrine transcriptional mechanism and endocytosis of extracellular galectin-7

doi: 10.1371/journal.pone.0187194

Figure Lengend Snippet: ( A ) Western blot analysis of cell lysates collected from MDA-MB-231 cells treated with rh-Gal-7 or FITC-rhGal-7 (15 min. post-treatment) ( B ) Expression of cytosolic or mitochondrial galectin-7 following treatment with rhGal-7 or FITC-rhGal-7 (15 min. post-treatment).

Article Snippet: Anti-human galectin-7 was purchased from R&D Systems (Minneapolis, MN, USA) while anti-lamin A/C, anti-Cox IV and anti-β-tubulin were obtained from Cell Signaling (Danvers, MA, USA).

Techniques: Western Blot, Expressing