Journal: Transplant international : official journal of the European Society for Organ Transplantation

Article Title: Intracellular signaling pathways control mitochondrial events associated with the development of ischemia/ reperfusion-associated damage.

doi: 10.1111/j.1432-2277.2009.00883.x

Figure Lengend Snippet: Figure 1 Alterations in MAPK activity are associated with IR or HR. Representative immunoblots (a, c, e) and a summary graph (b, d) are shown. (a, b). Three animals undergoing heterotopic heart transplantation were analysed in each group. Treatment conditions were as follows: Co, untreated animals; H, 45 min of ischemia; R1-R4, hearts exposed to 45 min of ischemia and either 10 min (R1), 2 h (R2), 12 h (R3), 24 h (R4) or 48 h (R5), of reperfusion. BALB/c mouse heart lysates display low levels of ERK1,2 activity in the control group (Co). 45 min of ischemia increased stress kinase activity (p38: 4.6-fold increase, *P < 0.05 vs. control; JNK: 9.4-fold increase, *P < 0.05 vs. control) but left ERK 1,2 unaf- fected. 10 min of reperfusion (R1) caused a significant increase in ERK activity (ERK1,2: 8.4-fold increase, **P < 0.01 vs. control), and a further increase in p38 (p38: 9.4-fold increase, **P < 0.01 vs. control) and JNK (JNK1,2: 57.7-fold increase, **P < 0.01 vs. control) activity. After 2 h of reperfusion (R2) JNK (JNK1,2: 2.6-fold increase) and p38 (p38: 0.6-fold) activity had returned to control (Co) levels, while ERK (ERK1,2: 4.8-fold increase) activation still did not return to prereperfusion levels and stayed above levels in control hearts until the end of the observation period (ERK1,2: 2.73-fold increase). Data are expressed as mean ± SEM (of n = 3). (c–e) Changes in MAPK signaling in HL-1 cells and primary BALB/c car- diomyocytes subjected to 45 min of hypoxia (0.5% O2, 37 C, serum-/glucose-free medium) and up to 48 h of reoxygenation. The treatment conditions were as follows: Co, untreated cells; H, 45 min of hypoxia R1-R4: hypoxia (45 min) followed by reoxygenation in growth medium for 10 min (R1), 2 h (R2), 24 h (R3) or 48 h (R4), respectively. 45 min of hypoxia increased activity of all three MAPKs, 10 min of reperfusion lead to a further increase in their activities. After 2 h of reperfusion JNK and p38 activity had ceased while ERK activity still had not returned to control levels. Data are expressed as mean ± SEM (n = 3, all **P < 0.01). In the case of primary cardiomyocytes a single experiment was performed.

Article Snippet: Immunoblots were probed with the following antibodies: phospho-ERK (sc-7383; Santa Cruz Biotechnology, Santa Cruz, CA, USA), phospho JNK (AF-1205; R&D Systems, Minneapolis, MN, USA) or phospho-p38 (9211S, Cell Signaling, Danvers, MA, USA).

Techniques: Activity Assay, Western Blot, Transplantation Assay, Control, Activation Assay

Journal: International Journal of Molecular Sciences

Article Title: Somatostatin, Cortistatin and Their Receptors Exert Antitumor Actions in Androgen-Independent Prostate Cancer Cells: Critical Role of Endogenous Cortistatin

doi: 10.3390/ijms232113003

Figure Lengend Snippet: Molecular consequences of somatostatin (SST) or cortistatin (CORT) treatment in androgen-independent (AI) prostate cancer (PCa) cells. ( A , B ) Phosphorylation levels of protein belonging to different oncogenic signaling pathways (AKT, ERK, JNK, PTEN and AR) in response to SST and CORT treatment in AI-PCa cells. Phospho-protein levels were normalized by the total amount of each respective protein. Protein data were represented as percent of vehicle-treated cells (set at 100%). ( C ) Fold change in markers of proliferation, migration, and PCa-aggressiveness in response to SST and CORT treatment in AI-PCa cells. Gene expression was represented as the percentage of vehicle-treated cells (set at 100%). Asterisks (* p < 0.05; ** p < 0.01 and *** p < 0.001) indicate statistically significant differences between treatment and vehicle-treated cells. N.D: Non-detected. Ctrl: Control.

Article Snippet: Membranes were blocked with 5% non-fat dry milk in Tris-buffered saline/0.05% Tween-20 and incubated overnight with the specific primary antibodies at 1:1000 dilution [phospho-AKT (p-AKT; #4060S; Cell-Signaling, Barcelona, Spain), AKT (#9272S; Cell-Signaling, Barcelona, Spain), phospho-ERK (#4370S; Cell-Signaling), ERK (#9102S; Cell-Signaling), phospho-JNK (#AF1206; RD system), JNK (#AF1387; RD system), phospho-PTEN (#S380; Cell-Signaling), PTEN (9552S; Cell-Signaling), phospho-AR (#16969; Cell-Signaling), AR (ab133273; Abcam)].

Techniques: Phospho-proteomics, Protein-Protein interactions, Migration, Gene Expression, Control

Journal: Frontiers in Immunology

Article Title: Platelet-rich plasma-derived microRNA let-7a-5p alleviates knee osteoarthritis by regulating macrophage polarization and improving inflammatory microenvironment

doi: 10.3389/fimmu.2026.1756467

Figure Lengend Snippet: Validation of the targeting relationship between let-7a-5p and MAPK8. (A) Schematic representation of the predicted complementary binding site between let-7a-5p and the 3′-UTR of MAPK8. (B) Relative mRNA expression level of MAPK8 in macrophages overexpressing let-7a-5p detected by RT-qPCR. (C) WB analysis of MAPK8 protein expression in macrophages overexpressing let-7a-5p. (D) Inhibitory effect of let-7a-5p on MAPK8 expression was assessed via a dual luciferase reporter assay. ( **P < 0.01, ***P < 0.001).

Article Snippet: Subsequently, the tissue sections and cells were incubated overnight at 4°C with primary antibodies against iNOS (Proteintech 22226-1-AP), CD206 (Proteintech 18704-1-AP) and MAPK8 (Proteintech 66210-1-Ig).

Techniques: Biomarker Discovery, Binding Assay, Expressing, Quantitative RT-PCR, Luciferase, Reporter Assay

Journal: Frontiers in Immunology

Article Title: Platelet-rich plasma-derived microRNA let-7a-5p alleviates knee osteoarthritis by regulating macrophage polarization and improving inflammatory microenvironment

doi: 10.3389/fimmu.2026.1756467

Figure Lengend Snippet: Effect of PRP on expression of let-7a-5p and MAPK8. (A) Relative mRNA expression levels of let-7a-5p in knee joint sections of each group detected by RT-qPCR. (B) iNOS and CD206 co-immunolabeld with MAPK8 and counter-stained with DAPI in synovial tissues (Scale bar: 50 μm). (C, D) Quantification of iNOS and CD206 expression co-localized with MAPK8. ( **P < 0.01).

Article Snippet: Subsequently, the tissue sections and cells were incubated overnight at 4°C with primary antibodies against iNOS (Proteintech 22226-1-AP), CD206 (Proteintech 18704-1-AP) and MAPK8 (Proteintech 66210-1-Ig).

Techniques: Expressing, Quantitative RT-PCR, Staining

Journal: Frontiers in Immunology

Article Title: Platelet-rich plasma-derived microRNA let-7a-5p alleviates knee osteoarthritis by regulating macrophage polarization and improving inflammatory microenvironment

doi: 10.3389/fimmu.2026.1756467

Figure Lengend Snippet: The let-7a-5p/MAPK8 axis regulates macrophage polarization and inflammatory cytokine release in vitro . (A) IF staining showing expression of iNOS and CD206 in macrophages (Scale bar: 50 μm). (B) Quantification of iNOS-positive cell rate in transfected macrophages. (C) Quantification of CD206-positive cell rate in transfected macrophages. (D-G) Relative mRNA expression levels of pro-inflammatory cytokine (IL-1β and TNF-α) and anti-inflammatory cytokine (IL-4 and IL-10) in transfected macrophages detected by RT-qPCR. ( *P < 0.05, **P < 0.01, ns no significance).

Article Snippet: Subsequently, the tissue sections and cells were incubated overnight at 4°C with primary antibodies against iNOS (Proteintech 22226-1-AP), CD206 (Proteintech 18704-1-AP) and MAPK8 (Proteintech 66210-1-Ig).

Techniques: In Vitro, Staining, Expressing, Transfection, Quantitative RT-PCR

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