Journal: The Journal of Physiological Sciences : JPS

Article Title: Identification of a small molecule that targets activator of G-protein signaling 8

doi: 10.1016/j.jphyss.2026.100064

Figure Lengend Snippet: Effects of AGS8 inhibitor on interaction assay and VEGF signaling in cells. A, Example of a GST pulldown assay of GST-AGS8 with Gβ 1 γ 2 . GST-fusion protein (10 pmol) and human Gβ 1 γ 2 (100 pmol) were incubated in 500 µL buffer for 16 h at 4℃ in the presence of AGS8 inhibitor. Five uL of mixtures were saved as “input”, then 20 µL of 50 % slurry of Glutathione Sepharose 4B was added. After 3 h incubation at 4°C, pellets were washed three times with 1000 µL buffer. This is a representative result from 4 independent experiments with similar results. The left panel and the right panels were captured from the same blot (see ). B, Effects of AGS8 inhibitor on VEGF-induced phosphorylation of VEGFR2, p38 MAPK and ERK1/2. Cell lysates (10–20 μg) from HUVECs treated with VEGF (10 ng/ml) for 10 min were subjected to immunoblotting, and signal intensity was quantified by densitometric analysis. In some groups, cells were treated with AGS8 inhibitor 30 min before stimulation. Values were normalized to total VEGFR2, p38 MAPK, and ERK1/2, respectively. Data shown are representative pictures from 5 independent experiments with similar results (n = 5). P < 0.05, control vs. VEGF (two-way ANOVA) in each of VEGFR2, p38 MAPK, and ERK1/2. ＊ P < 0.05 vs . VEGF without AGS8 inhibitor (one-way ANOVA with Dunnett’s multiple-comparison test).

Article Snippet: Antibodies against VEGFR2 (#2479), phospho-VEGFR2 (Tyr1175) (#3770), p38MAPK family proteins (#9212), phospho-p38 MAPK family proteins (Thr180/Tyr182) (#9211), ERK1/2 (p44/42 MAPK) (#9102), and phospho-ERK1/2 (p44/42 MAPK) (Thr 202/204) (#9101) were purchased from Cell Signaling Technology (Danvers, MA).

Techniques: GST Pulldown Assay, Incubation, Phospho-proteomics, Western Blot, Control, Comparison

Journal: Cancer Medicine

Article Title: Carnosic Acid Mediates Production of Reactive Oxygen Species to Regulate Mitogen‐Activated Protein Kinase Pathway Phosphorylation and Induce Apoptosis in Human Breast Cancer Cells

doi: 10.1002/cam4.71446

Figure Lengend Snippet: CA induced breast cancer cell apoptosis via p38 and JNK phosphorylation and regulated glucose metabolism. (A) T‐47D and MCF‐7 cells were treated with 25 μM and 20 μM CA for 6, 12, and 24 h, respectively. Cancer cell apoptosis was detected by flow cytometry using annexin V‐FITC/PI (B) Potential targets of CA. (C) Protein expression levels of p‐JNK, p‐p38, Bax, Bcl‐2, and PARP were measured by western blotting. Data presented as mean ± SD of three independent experiments. * p < 0.05, ** p < 0.01 versus NC group.

Article Snippet: The following antibodies were also used: rabbit monoclonal phospho (p)‐JNK, p‐p38 (Cell Signaling Technology, 9926 T), rabbit polyclonal β‐actin (Proteintech, 20,536–1‐AP), Bax (Proteintech, 50,599‐2‐Ig), Bcl‐2 (Proteintech, 12,789‐1‐AP), and poly‐ADP ribose polymerase (PARP, Cell Signaling Technology, 9542).

Techniques: Phospho-proteomics, Flow Cytometry, Expressing, Western Blot

Journal: Cancer Medicine

Article Title: Carnosic Acid Mediates Production of Reactive Oxygen Species to Regulate Mitogen‐Activated Protein Kinase Pathway Phosphorylation and Induce Apoptosis in Human Breast Cancer Cells

doi: 10.1002/cam4.71446

Figure Lengend Snippet: SP600125 and BMS582949 were used to inhibit CA‐induced JNK and p38 phosphorylation. (A) T‐47D and (B) MCF‐7 cells were pre‐treated with SP600125 and BMS582949 for 2 h, followed by treatment with CA at 25 μM and 20 μM, respectively. Protein expression levels of p‐JNK, p‐p38, Bax, Bcl‐2, and PARP were measured by western blotting. (C) and (D) Relative cell viability was evaluated by MTT assays. Data are presented as mean ± SD of three independent experiments. * p < 0.05, ** p < 0.01 versus NC group.

Article Snippet: The following antibodies were also used: rabbit monoclonal phospho (p)‐JNK, p‐p38 (Cell Signaling Technology, 9926 T), rabbit polyclonal β‐actin (Proteintech, 20,536–1‐AP), Bax (Proteintech, 50,599‐2‐Ig), Bcl‐2 (Proteintech, 12,789‐1‐AP), and poly‐ADP ribose polymerase (PARP, Cell Signaling Technology, 9542).

Techniques: Phospho-proteomics, Expressing, Western Blot