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: 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