Journal: Scientific Reports

Article Title: Aerobic exercise promotes the expression of ATGL and attenuates inflammation to improve hepatic steatosis via lncRNA SRA

doi: 10.1038/s41598-022-09174-0

Figure Lengend Snippet: Aerobic exercise improves P38/JNK-mediated inflammation through SRA. ( A ) The mRNA levels of inflammatory factors in mice liver assessed by RT-qPCR, n = 6/group. ( B ) Western blot analysis of protein expression and phosphorylation of P38 and JNK. ( C ) The phosphorylated P38/total protein levels in the liver of each group of mice. ( D ) The Phosphorylated JNK/total protein levels in the liver of each group of mice. a P < 0.05 vs. NC group; b P < 0.05 vs. HFD group; c P < 0.05 vs. NC group. ( E ) The mRNA levels of inflammatory factors in the liver of EV, SRA+, SRA + AE mice were assessed by RT-qPCR, n = 6/group. ( F ) Western blot analysis of protein expression and phosphorylation of P38 and JNK in EV, SRA+, SRA + AE mice. ( G ) The phosphorylated P38/total protein levels in the liver of each group of mice. ( H ) The Phosphorylated JNK/total protein levels in the liver of each group of mice. (I) The expression of PERK activity in mice liver, n = 6/group. ( J ) The expression of IRE1a activity in mice liver, n = 6/group. a P < 0.05 vs. NC group; b P < 0.05 vs. HFD group; * P < 0.05 vs. EV group; # P < 0.05 vs. SRA + group. The values represent the means ± SD. Abbreviations: NC, normal control group; NE, normal aerobic exercise group; HFD, high-fat diet group and HAE, high-fat diet with aerobic exercise group; EV virus empty vector group; SRA+, SRA overexpression group; SRA + AE, SRA overexpression and combined with the aerobic exercise group.

Article Snippet: After overnight incubation with FoxO1 (1:1000, 18592-1-AP, Wuhan Sanying Biotechnology Co., Ltd.), P-FoxO1 (1:1000, ab131339, Abcam), ATGL (1:1000, 55190-1-AP, Wuhan Sanying Biotechnology Co., Ltd.), P38 (1:1000, 8690 T, CST), P-P38 (1:1000, 9211S, CST), JNK (1:1000, 24164-1-AP, Wuhan Sanying Biotechnology Co., Ltd.), P-JNK (1:1000, 24164-1-AP, Wuhan Sanying Biotechnology Co., Ltd.), P-P38 (1:1000, 9211S, CST), JNK (1:1000, 24164-1-AP, Wuhan Sanying Biotechnology Co., Ltd.), P-JNK (1:2000, Sc-6254, Santa Cruz), and β-actin (1:500, BM0627, Wuhan PhD Bioengineering Co., Ltd.) primary antibody.

Techniques: Quantitative RT-PCR, Western Blot, Expressing, Phospho-proteomics, Activity Assay, Control, Virus, Plasmid Preparation, Over Expression

Journal: Scientific Reports

Article Title: Aerobic exercise promotes the expression of ATGL and attenuates inflammation to improve hepatic steatosis via lncRNA SRA

doi: 10.1038/s41598-022-09174-0

Figure Lengend Snippet: Aerobic exercise via SRA improves hepatic steatosis description model. Aerobic exercise is closely linked to the reduction of intrahepatic lipid aggregation and inflammatory responses through the inhibition of SRA levels. Mechanistically, aerobic exercise may inhibit the transcriptional activity of FoxO1 by suppressing SRA expression, which leads to upregulation of ATGL expression. In addition, remission of inflammation was accompanied by significant alteration of inflammatory proteins and the P38/JNK signaling pathway.

Article Snippet: After overnight incubation with FoxO1 (1:1000, 18592-1-AP, Wuhan Sanying Biotechnology Co., Ltd.), P-FoxO1 (1:1000, ab131339, Abcam), ATGL (1:1000, 55190-1-AP, Wuhan Sanying Biotechnology Co., Ltd.), P38 (1:1000, 8690 T, CST), P-P38 (1:1000, 9211S, CST), JNK (1:1000, 24164-1-AP, Wuhan Sanying Biotechnology Co., Ltd.), P-JNK (1:1000, 24164-1-AP, Wuhan Sanying Biotechnology Co., Ltd.), P-P38 (1:1000, 9211S, CST), JNK (1:1000, 24164-1-AP, Wuhan Sanying Biotechnology Co., Ltd.), P-JNK (1:2000, Sc-6254, Santa Cruz), and β-actin (1:500, BM0627, Wuhan PhD Bioengineering Co., Ltd.) primary antibody.

Techniques: Inhibition, Activity Assay, Expressing

Journal: International Journal of Molecular Sciences

Article Title: Kappa-Opioid Receptor Antagonism Prolongs the Antidepressant Effects of Ketamine in Adult Mice with Depression-like Behavior Induced by Adolescent Chronic Unpredictable Stress

doi: 10.3390/ijms27062815

Figure Lengend Snippet: Effects of CUS and treatments on ERK, AKT, JNK, and mTOR activation in the PFC. ( A ) Phosphorylation ratio of ERK (p-ERK/ERK). ( B ) Phosphorylation ratio of AKT (p-AKT/AKT). ( C ) Phosphorylation ratio of JNK (p-JNK/JNK). ( D ) Phosphorylation ratio of mTOR (p-mTOR/mTOR). Differences among all groups were analyzed by one-way ANOVA, with Tukey post hoc comparisons made against the CUS group. n = 4–10. Data are expressed as mean ± SEM. * p < 0.05, ** p < 0.01, **** p < 0.0001. Abbreviations: CUS, chronic unpredictable stress; Ket, ketamine; nBNI, norbinaltorphimine; Ket/nBNI, combination treatment with ketamine and norbinaltorphimine.

Article Snippet: p-JNK (Thr 183/Tyr 185) , 1:500 , Santa Cruz Biotechnology , sc-6254.

Techniques: Activation Assay, Phospho-proteomics

Journal: International Journal of Molecular Sciences

Article Title: Kappa-Opioid Receptor Antagonism Prolongs the Antidepressant Effects of Ketamine in Adult Mice with Depression-like Behavior Induced by Adolescent Chronic Unpredictable Stress

doi: 10.3390/ijms27062815

Figure Lengend Snippet: Effects of CUS and treatments on ERK, AKT, JNK, and mTOR activation in the hippocampus. ( A ) Phosphorylation ratio of ERK (p-ERK/ERK). ( B ) Phosphorylation ratio of AKT (p-AKT/AKT). ( C ) Phosphorylation ratio of JNK (p-JNK/JNK). ( D ) Phosphorylation ratio of mTOR (p-mTOR/mTOR). n = 4–10. Differences among all groups were analyzed by one-way ANOVA, with post hoc comparisons made against the CUS group. Data are expressed as mean ± SEM. * p < 0.05, ** p < 0.01, **** p < 0.0001, ns, not significant.

Article Snippet: p-JNK (Thr 183/Tyr 185) , 1:500 , Santa Cruz Biotechnology , sc-6254.

Techniques: Activation Assay, Phospho-proteomics

Journal: International Journal of Molecular Sciences

Article Title: Kappa-Opioid Receptor Antagonism Prolongs the Antidepressant Effects of Ketamine in Adult Mice with Depression-like Behavior Induced by Adolescent Chronic Unpredictable Stress

doi: 10.3390/ijms27062815

Figure Lengend Snippet: Effects of CUS and treatments on ERK, AKT, JNK, and mTOR activation in the striatum. ( A ) Phosphorylation ratio of ERK (p-ERK/ERK). ( B ) Phosphorylation ratio of AKT (p-AKT/AKT). ( C ) Phosphorylation ratio of JNK (p-JNK/JNK). ( D ) Phosphorylation ratio of mTOR (p-mTOR/mTOR). n = 4–10. Differences among all groups were analyzed by one-way ANOVA, with post hoc comparisons made against the CUS group. Data are expressed as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: p-JNK (Thr 183/Tyr 185) , 1:500 , Santa Cruz Biotechnology , sc-6254.

Techniques: Activation Assay, Phospho-proteomics

Journal: Cell communication and signaling : CCS

Article Title: Matrix stiffness-induced IKBKE and MAPK8 signaling drives a phenotypic switch from DCIS to invasive breast cancer.

doi: 10.1186/s12964-025-02276-y

Figure Lengend Snippet: Fig. 3 MAPK8 and IKBKE mediate the stiffness-induced breast cancer phenotype. a, Venn diagram (left) and table (right) showing overlap between ki nases predicted to be upregulated by stiffness and downregulated by AIIB2 treatment. b, Workflow for the siRNA-based screen used to test the function of predicted kinases, and all conditions were run in duplicates. c, Representative phenotype of CA1a cells on high stiffness hydrogels transfected with siRNA as specified. FAK siRNA serves as positive control. Images are single tiles from 6 × 6 montages. Scale bar = 50 μm. d, Area fraction of segmented cell clusters expressed relative to MOCK (Gene of interest/MOCK). The area fraction is calculated as the ratio between the total area and the area of the thresholded objects. Positive control (FAK siRNA) is shown in red. Data represents the average of two technical replicates. Genes whose knockdown results in a similar or lower relative area fraction as the positive control are considered hits, including MAPK8 and IKBKE

Article Snippet: Membranes were blocked with 5% non-fat dry milk (PanReac AppliChem) in TBST (20 mM Tris, 150 mM NaCl, 0.1% Tween-20, pH 7.4) for 1 h at RT, followed by overnight incubation at 4 °C with primary antibodies against IKBKE (Cell Signaling Technology, Cat. #D20G4, 1:1000), phosphorylated-MAPK8 (Novus Biologicals; Cat. #NB100-82009; 1:2000), or total-MAPK8 (Santa Cruz; Cat. #sc-1648; 1:5000).

Techniques: Transfection, Positive Control, Knockdown

Journal: Cell communication and signaling : CCS

Article Title: Matrix stiffness-induced IKBKE and MAPK8 signaling drives a phenotypic switch from DCIS to invasive breast cancer.

doi: 10.1186/s12964-025-02276-y

Figure Lengend Snippet: Fig. 5 Matrix stiffness-induced MAPK8 activity is critical for breast cancer cell proliferation in vitro. a, Immunoblot analysis of MAPK8 (left), and phospho- MAPK8 (right) of CA1a cells cultured on 0.4 kPa or 5 kPa hydrogels. Densitometric analysis shows MAPK8 and phospho-MAPK8 levels normalized to load ing controls (ponceau S staining, Supplementary Fig. 2a and 2c) and expressed relative to the low stiffness levels. Data are presented as mean ± S.E.M., with p-values according to an unpaired t-test. b, Representative Edu staining images of CA1a cells grown on 5 kPa transfected with MAPK8 or control siRNA pool (left) and quantification of three biological repeats (> 1000 cells each; right). Scale bar = 50 μm. Data are mean ± S.E.M and p-values according to an unpaired t-test. c, Representative images of actin (top row) and Edu staining (bottom row) in HCC1143 cells treated with MAPK8 or control siRNAs (left). Quantification of area fraction of segmented cell clusters relative to control siRNA of three biological repeats (right). Scale bar = 50 μm. Data are mean ± S.E.M and p-values derived by one-way Anova with Dunnett’s multiple comparison test. d, e, Spinning disc confocal images of breast cancer cells on 5 kPa treated with 5 µM (CA1a, d) or 10 µM ( HCC1143, e) JNK-IN-8 or vehicle for 24 h(left). Quantification of actin staining (top row) and Edu staining (bottom row) of three or more independent experiments(right). Scale bar = 50 μm. Relative area fraction as in (c). Data and statistics as in (b)

Article Snippet: Membranes were blocked with 5% non-fat dry milk (PanReac AppliChem) in TBST (20 mM Tris, 150 mM NaCl, 0.1% Tween-20, pH 7.4) for 1 h at RT, followed by overnight incubation at 4 °C with primary antibodies against IKBKE (Cell Signaling Technology, Cat. #D20G4, 1:1000), phosphorylated-MAPK8 (Novus Biologicals; Cat. #NB100-82009; 1:2000), or total-MAPK8 (Santa Cruz; Cat. #sc-1648; 1:5000).

Techniques: Activity Assay, In Vitro, Western Blot, Cell Culture, Staining, Transfection, Control, Derivative Assay, Comparison