Journal: Oncogene

Article Title: Copper ionophore elesclomol selectively targets GNAQ/11-mutant uveal melanoma.

doi: 10.1038/s41388-022-02364-0

Figure Lengend Snippet: Fig. 6 Elesclomol promotes YAP phosphorylation via ROS-induced Hippo signaling pathway activation. A YAP protein and its phosphorylation level were detected via western blotting in tumor samples from eight UM patients. B The p-YAP/YAP ratio was calculated and compared between GNAQ/11-mutant and GNAQ/11-wild-type primary tumor tissues. C Representative dose–response curves for elesclomol- treated OMM2.3 cells transfected with siYAP or control siRNA. D Representative dose–response curves for elesclomol-treated 92.1 cells transfected with YAPS127A-Flag or control plasmid. E Immunoblot analysis and quantification of p-LATS1 (phospho Thr1079), total LATS1, p- MST1/MST2 (phospho Thr180 + Thr183) and total MST1 in UM cells treated with elesclomol at select concentrations for 3 h. GAPDH was used as the loading control. F Immunoblot analysis and quantification of p-LATS1, LATS1, p-YAP, and YAP in UM cells treated with elesclomol with/ without NAC or rotenone for 3 h. GAPDH was used as the loading control. Error bars represent the mean ± SEM, n = 3 independent experiments for (A, C, D, E, F), *p < 0.05; **p < 0.01; ***p < 0.001.

Article Snippet: Primary antibodies targeting the following proteins were used: c-Myc (Abcam, ab32072), cyclin D1 (Abcam, ab134175), caspase3 (Proteintech, 19677-1-AP), cleaved caspase-3 (Cell Signaling, 9661), p-ERK1/2 (phospho Thr202/Tyr204, Cell Signaling, 4370), ERK1/2 (Cell Signaling, 9107), p-YAP (phospho Ser127, Abcam, ab76252), YAP (Proteintech, 66900-1-Ig), p-AKT (phospho Thr308, Cell Signaling, 13038), AKT (Cell Signaling, 4691), p-LATS1 (phospho Thr1079, Cell Signaling, 8654), LATS1 (Proteintech, 17049-1-AP), p-MST1/MST2 (phospho Thr180+ Thr183, Abcam, ab76323), MST1 (Proteintech, 22245-1-AP), Flag (Cell Signaling, #14793), SNAI2 (Proteintech, 12129-1-AP), GNAQ (Proteintech, 13927-1-AP), and GAPDH (Abcam, ab8245).

Techniques: Phospho-proteomics, Activation Assay, Western Blot, Mutagenesis, Transfection, Control, Plasmid Preparation

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: STAT1 and STAT6 orchestrate Cbs transcription and transsulfur metabolism in microglia and contribute to parkinson’s disease-related neuroinflammation

doi: 10.1007/s00018-025-05768-9

Figure Lengend Snippet: CBS expression was upregulated during microglial anti-inflammatory activation triggered by IL-4 and IL-13. ( A , B ) qPCR analysis of Cd206 and Cbs mRNA levels in BV2 cells treated with IL-4 (20 ng/ml) + IL-13 (20 ng/ml) for 0, 6, 12 and 24 h; n = 3. ** P ˂0.01, *** P ˂0.001 vs. the 0 h group. ( C , D ) Western blot data for ARG1, CBS, CSE and 3-MST in IL-4 + IL-13–treated BV2 cells at various time points, n = 3. * P ˂0.05, ** P ˂0.01, *** P ˂0.001 vs. the 0 h group. One-way ANOVA followed by Dunnett’s post hoc analysis. ( E–G ) Western blot analysis of ARG1 and CBS protein levels in si-Cbs- or si-NC- transfected BV2 cells following IL-4 + IL-13 treatment; n = 3. ** P ˂0.01 vs. the si-NC group; ### P ˂0.001 vs. the si-NC with IL-4 + IL-13 group. ( H ) qPCR data of Arg1 and Cb206 mRNA levels in WT and Cbs −/− microglia following IL-4 + IL-13 treatment, n = 3. ** P ˂0.01, *** P ˂0.001 vs. the IL-4 + IL-13-treated WT group. Two-way ANOVA followed by Tukey’s post hoc analysis for F‒H

Article Snippet: Antibodies against 3-mercaptopyruvate sulfurtransferase (3-MST) (A11587), p-STAT1 Y701 and STAT1 (AP0054/A19563) were obtained from ABclonal (China).

Techniques: Expressing, Activation Assay, Western Blot, Transfection

Journal: Current Issues in Molecular Biology

Article Title: Levistolide A Alleviates Myocardial Ischemia–Reperfusion Injury Partly by Improving Calcium Homeostasis via the ADORA2B/cAMP/PKA/PLB/SERCA2α Signaling Axis

doi: 10.3390/cimb48020125

Figure Lengend Snippet: The key role of ADORA2B in the calcium signaling pathway and its molecular docking results with LA. ( A ) Schematic representation of the central position of ADORA2B in the calcium signaling pathway; ( B ) Molecular docking binding mode of LA with the ADORA2B protein.

Article Snippet: The antibodies used in this study are as follows: adenosine A2B receptor (ADORA2B) (37KDa) antibody (bs-5900R) was purchased from Bioss Biotechnology Co., Ltd. (Beijing, China); phospholamban (PLB) (36KDa) antibody (A01395-1) was purchased from Boster Biological Engineering Co., Ltd. (Wuhan, China); phosphorylated phospholamban-Ser16 (p-PLB-Ser16) (36KDa) antibody (AP0907) was purchased from ABclonal Biotechnology Co., Ltd. (Wuhan, China); SERCA2α (140KDa) antibody (9580T) was purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA); and α-Tubulin (55 KDa) antibody (66031-1-Ig) was purchased from Proteintech Group, Inc. (Wuhan, China).

Techniques: Binding Assay

Journal: Current Issues in Molecular Biology

Article Title: Levistolide A Alleviates Myocardial Ischemia–Reperfusion Injury Partly by Improving Calcium Homeostasis via the ADORA2B/cAMP/PKA/PLB/SERCA2α Signaling Axis

doi: 10.3390/cimb48020125

Figure Lengend Snippet: Protein expression levels of ADORA2B, p-PLB, PLB, and SERCA2α in different groups of H9C2 cells. ( A ) Representative Western blot bands of target proteins in each group; ( B ) Relative protein expression level of ADORA2B; ( C ) Relative protein expression level of p-PLB; ( D ) Relative protein expression level of SERCA2α. Compared to the control group, ## p < 0.01. Compared to the H/R group, ** p < 0.01. Compared to the LA + H/R group, Δ p < 0.05, ΔΔ p < 0.01.

Article Snippet: The antibodies used in this study are as follows: adenosine A2B receptor (ADORA2B) (37KDa) antibody (bs-5900R) was purchased from Bioss Biotechnology Co., Ltd. (Beijing, China); phospholamban (PLB) (36KDa) antibody (A01395-1) was purchased from Boster Biological Engineering Co., Ltd. (Wuhan, China); phosphorylated phospholamban-Ser16 (p-PLB-Ser16) (36KDa) antibody (AP0907) was purchased from ABclonal Biotechnology Co., Ltd. (Wuhan, China); SERCA2α (140KDa) antibody (9580T) was purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA); and α-Tubulin (55 KDa) antibody (66031-1-Ig) was purchased from Proteintech Group, Inc. (Wuhan, China).

Techniques: Expressing, Western Blot, Control