Journal: International Journal of Biological Sciences

Article Title: TREM-1 governs NLRP3 inflammasome activation of macrophages by firing up glycolysis in acute lung injury

doi: 10.7150/ijbs.77304

Figure Lengend Snippet: Antibody sources and dilutions

Article Snippet: Anti-mTOR monoclonal antibody , Proteintech , 66888-1-lg , 1:2000.

Techniques:

Journal: International Journal of Biological Sciences

Article Title: TREM-1 governs NLRP3 inflammasome activation of macrophages by firing up glycolysis in acute lung injury

doi: 10.7150/ijbs.77304

Figure Lengend Snippet: Blockade of TREM-1 reduced intrapulmonary inflammation and limited glycolysis in LPS-induced ALI mice. C57BL/6J mice were intravenously injected with LR12 (5 mg/kg) 2 h before the LPS administration (5 mg/kg, i.t. ). (A) Six hours later, mouse lungs were excised, and lung histopathology was performed with H&E staining. One representative picture of six mice in each group is shown. (B) Inflammation score was measured, n =6 mice/group. (C) Pro-IL-1β, IL-1β p17, and iNOS in the lung lysates were assessed by western blot with β-actin as a loading control. (D-F) The western blot results were quantitated using Image Lab, n =8 mice per group. (G-H) Lactate concentration in BALF and serum was assayed, n =4-9 mice/group. (I) Expression of Hk2 , Pfkfb3, and Hif-1α mRNA in the lungs was detected by real-time PCR. Data was normalized to housekeeping gene β-actin, n =5-10 mice/group. (J) Glycolysis-associated proteins of HK2, p-mTOR, mTOR, and HIF-1α in the lung lysates were assessed by western blot with α-tubulin as a loading control. (K-N) Quantification of indicated protein levels in (J), n =6-8 mice/group. In all cases, the experiment was repeated twice. Dots represent individual animal values. Statistical analysis was performed using One-way ANOVA adjusted by Tukey's multiple comparison test for Control vs. ALI or ALI vs. LR12+ALI. Error bars indicate mean ± SD. * P < 0.05, ** P < 0.01, and *** P < 0.001. Original western blots represented in graphs are available in Figure -source data.

Article Snippet: Anti-mTOR monoclonal antibody , Proteintech , 66888-1-lg , 1:2000.

Techniques: Injection, Histopathology, Staining, Western Blot, Control, Concentration Assay, Expressing, Real-time Polymerase Chain Reaction, Comparison

Journal: International Journal of Biological Sciences

Article Title: TREM-1 governs NLRP3 inflammasome activation of macrophages by firing up glycolysis in acute lung injury

doi: 10.7150/ijbs.77304

Figure Lengend Snippet: TREM-1 activation stimulated HIF-1α accumulation via PI3K/AKT/mTOR signaling. 1×10 6 macrophages/well were plated into 12-well plates with agonist anti-TREM-1 mAb (10 μg/mL). (A) p-PI3K p85α T607 , total PI3K, p-AKT s473 , AKT, p-mTOR s2448 , and mTOR protein levels in control or 24 h TREM-1-activated macrophages. (B-D) Quantification of p-PI3K p85α T607 , p-AKT s473 and p-mTOR s2448 in (A), n =3 biological replicates. Statistical analysis was performed using Student's t -test (two-tailed, unpaired). (E) Lactate secretion, (F) glucose consumption in the supernatant, and (G-H) HIF-1α protein levels in control or 24 h TREM-1-activated macrophages co-treated with or without LY294002 (25 μM). n =3 biological replicates. Statistical analysis was performed using One-way ANOVA. (I) Lactate secretion, (J) glucose consumption in the supernatant, and (K-L) HIF-1α protein levels in control or 24 h TREM-1-activated macrophages co-treated with or without Rapamycin (100 nM). n =3 biological replicates. Statistical analysis was performed using One-way ANOVA adjusted by Tukey's multiple comparison test. Data were expressed as the mean ± SD. * P < 0.05, ** P < 0.01, and *** P < 0.001.

Article Snippet: Anti-mTOR monoclonal antibody , Proteintech , 66888-1-lg , 1:2000.

Techniques: Activation Assay, Control, Two Tailed Test, Comparison

Journal: International Journal of Biological Sciences

Article Title: TREM-1 governs NLRP3 inflammasome activation of macrophages by firing up glycolysis in acute lung injury

doi: 10.7150/ijbs.77304

Figure Lengend Snippet: Schematic illustration. TREM-1 activation stimulates HIF-1α induced glucose metabolic reprogramming via PI3K/AKT/mTOR pathway. HIF-1α-induced glycolysis promotes TREM-1-governed NLRP3 inflammasome activation, facilitating intrapulmonary inflammation in ALI.

Article Snippet: Anti-mTOR monoclonal antibody , Proteintech , 66888-1-lg , 1:2000.

Techniques: Activation Assay

Journal: Neuropsychiatric Disease and Treatment

Article Title: Shisandra Decoction Alleviates Parkinson’s Disease Symptoms in a Mouse Model Through PI3K/AKT/mTOR Signalling Pathway

doi: 10.2147/NDT.S476969

Figure Lengend Snippet: AKT and mTOR phosphorylation and relative abundance of p70S6K. ( A ) WB electrophoretogram. ( B ) and ( C ) Relative degree of AKT and mTOR phosphorylation. ( D ) Relative abundance of p70S6K. Use the Tukey HSD test statistical method. *P < 0.05, **P < 0.01 compared to control. # P < 0.05, ## P < 0.01 compared to model group. Δ P < 0.05, ΔΔ P < 0.01 compared to LY294002 group.

Article Snippet: MPTP (Shanghai Yuanye Biotechnology Co., Ltd., item number S31504-500mg); LY294002 (MedChemExpress, item number HY-10108); LC3 antibody, AKT antibody, p-AKT antibody, PTEN antibody, PI3K antibody, mTOR antibody, p-mTOR antibody, p70S6K antibody, p62/SQSTM1 antibody, α-syn antibody (Wuhan Biode Biotechnology Engineering Co., Ltd., item numbers: BM4827, BM1612, BM4762, A00006-1, PB0351, BM4182, BM4840, M01475-4, PB0458, M00215-3, respectively).

Techniques: Phospho-proteomics, Control

Journal: Cell Death & Disease

Article Title: Microglia as modulators of exosomal alpha-synuclein transmission

doi: 10.1038/s41419-019-1404-9

Figure Lengend Snippet: Whole-cell lysates were analyzed by western blot. a Protein levels of LC3 ( n = 5, ** p = 0.0082), P62/SQSTM1 ( n = 5, **** p < 0.0001), Beclin ( n = 5, * p = 0.0101), phospho-AKT ( n = 5, *** p = 0.0004), total-AKT, phospho-mTOR ( n = 4, * p = 0.0183), and β-actin were shown in indicated cells. b Whole-cell lysates were immunoblotted against α-syn (oligomeric) ( n = 5, ** p = 0.0078); monomeric ( n = 4, * p = 0.0113) and β-actin for control. c The triple-stained image of P62/SQSTM1 (gray) with IBA1 (green) and DAPI (blue) in the SNpc

Article Snippet: The following primary antibodies were utilized: rabbit antibody to IBA1 (1:1000, 10904-1-AP, Proteintech), mouse antibody to tsg 101 (1:500, sc-7964, Santa cruz biotechnology), mouse antibody to CD63 (1:1000, ab59479, Abcam), rabbit antibody to Calnexin (1:1000, 10427-2-AP, Proteintech), rabbit antibody to α-syn (1:1000, ab138501, ab52168, Abcam), rabbit antibody to Oligomer (1:1000, AHB0052, ThermoFisher scientific), rabbit antibody to TH (1:1000, 25859-1-AP, Proteintech), rabbit antibody to LC3 (1:1000, 14600-1-AP, Proteintech), rabbit antibody to SQSTM1/P62 (1:1000, ab91526, Abcam), rabbit antibody to Beclin1 (1:1000, 11306-1-AP, Proteintech), rabbit antibody to phosphor-AKT (1:800, AP0655, Abclonal), rabbit antibody to AKT (1:800, A11030, Abclonal), rabbit antibody to phosphor-mTOR (1:400, BM4840, Boster), mouse antibody to α-syn (1:1000, synuclein Ab-2, Thermo Scientific), and rabbit antibody to p-syn (1:500, GTX50222, Genetex).

Techniques: Western Blot, Control, Staining

Journal: bioRxiv

Article Title: Argon inhibits autophagy and improves cerebral ischemia-reperfusion injury via PI3K/Akt/mTOR pathway

doi: 10.1101/2025.10.31.685964

Figure Lengend Snippet: A: western blot results showed that argon treatment significantly upregulated the levels of p-PI3K, p-Akt, and p-mTOR. B: Bar chart for A; C: western blot results of PI3K/Akt/mTOR pathway factors and autophagy factors in N 2, N 2 combined with SC79, and N 2 combined with MHY1485 group; D: Bar chart for C. E: western blot results of in argon, argon combined with MK2206 and argon combined with rapamycin groups. F: Bar chart for E. MHY: MHY1485; Rapa: rapamycin.

Article Snippet: The first antibodies included: P62 (Abcam, 1:1000, rabbit derived polyclonal antibody, ab91526), LC3 (Sigma Aldrich, 1:500, rabbit derived polyclonal antibody, ABC929), β-actin (MCE, 1:5000, mouse monoclonal antibody, HY-P80438), PI3K (Abcam, 1:1000, rabbit derived monoclonal antibody, ab302958), p-PI3K (Abcam, 1:1000, rabbit derived polyclonal antibody, ab138364), Akt (MCE, 1:2000, rabbit monoclonal antibody), p-Akt (MCE, 1:1000, rabbit monoclonal antibody, HY-P80276), mTOR (MCE, 1:1000, rabbit polyclonal antibody, HY-P80276), p-mTOR (MCE, 1:1000, rabbit monoclonal antibody, HY-P80837).

Techniques: Western Blot