Journal: Cell metabolism

Article Title: Targeting Peripheral CB 1 Receptors Reduces Ethanol Intake via a Gut-Brain Axis

doi: 10.1016/j.cmet.2019.04.012

Figure Lengend Snippet: (A) Time course and rate of increase in oleate-driven respiratory activity in MGN3–1 cells. Cells were treated with 10 or 100 nM JD5037, 2.5 M of FCCP (positive control) or vehicle in the presence of the fluorescent extracellular O2 consumption reagent. Oxidative respiration was recorded every 90 sec and the slope of the initial linear increase was calculated. Points and bars are means ± SEM from n = 11–15 experiments, as indicated. *P < 0.05 compared to vehicle. (B) Verification of Cnr1 knockdown by rtPCR in cells used in panel C. Cellular uptake of the constructs was verified by fluorescent microscopy (20x magnification) and the degree of knockdown was determined by rt-PCR, *P < 0.05, n = 4. (C) Oleate-driven oxidative activity in MGN3–1 cells with shRNA-mediated knockdown of Cnr1 expression and their mock-transfected controls, n = 3–8 experiments, as indicated, *P < 0.05.

Article Snippet: Scramble shRNA with GFP Adenovirus (Ad-scramble-shRNA) , Vector Biolabs , 1122.

Techniques: Activity Assay, Positive Control, Knockdown, Reverse Transcription Polymerase Chain Reaction, Construct, Microscopy, shRNA, Expressing, Transfection

Journal: Frontiers in Immunology

Article Title: The Crucial Role of PPARγ-Egr-1-Pro-Inflammatory Mediators Axis in IgG Immune Complex-Induced Acute Lung Injury

doi: 10.3389/fimmu.2021.634889

Figure Lengend Snippet: Ectopic expression of peroxisome proliferator-activated receptor (PPAR)γ in the lung inhibits IgG immune complex (IgG-IC)-induced acute lung injury. (A) Different doses of adenovirus are injected into lungs via airways. Seventy-two hours later, lungs are harvested and total proteins are extracted. Then Western blot assays are conducted by using antibodies recognizing PPARγ and GAPDH, respectively. Mice are treated by airway administration of 1 × 10 8 PFU of Ad-GFP or Ad-PPARγ. Three days later, acute lung injury is induced by treating the mice with IgG-IC. Four hours later, bronchoalveolar lavage (BAL) fluids and whole lungs are collected to analyze lung permeability indexes (B) , measurement of pulmonary myeloperoxidase (MPO) contents (C) , total white blood cells (D) , and neutrophils counts (E) in BAL fluids, and levels of TNF-α (F) , MCP-1 (G) , MIP-1α (H) , and MIP-2 (I) in BAL fluids, respectively. Data are expressed as means ± S. E. M. N=3 for α-BSA-treated mice, N=5 for Ad-GFP+IgG-IC group, and N=6 for Ad-PPARγ+IgG-IC group. *, ** and *** indicate statistically significant difference— p < 0.05, p < 0.01, and p < 0.001, respectively.

Article Snippet: Ad-GFP (BD Biosciences) or Ad-PPARγ is intratracheally administrated into mice.

Techniques: Expressing, Injection, Western Blot, Permeability

Journal: Frontiers in Immunology

Article Title: The Crucial Role of PPARγ-Egr-1-Pro-Inflammatory Mediators Axis in IgG Immune Complex-Induced Acute Lung Injury

doi: 10.3389/fimmu.2021.634889

Figure Lengend Snippet: IgG immune complex (IgG-IC)-mediated pulmonary early growth response protein 1 (Egr-1) expression is inhibited by ectopic production of peroxisome proliferator-activated receptor (PPAR)γ. Four hours after pulmonary deposition of IgG-IC, lungs are harvested. The tissue RNAs and proteins are isolated separately. Then the Egr-1 expression is examined by qPCR (A) and Western blot (B) , respectively. (C) Mice are treated with airway injection of Ad-GFP or Ad-PPARγ at a dose of 1 × 10 8 PFU. Seventy-two hours later, the mice are intratracheally challenged by IgG-IC for 4 h. Then the lungs are harvested, and proteins are extracted for Western blot analysis by using antibodies against Egr-1 and GAPDH, respectively. Data are expressed as means ± S. E. M. (For qPCR, N=3 for α-BSA treated mice, and N=5 for IgG-IC-treated mice; N=3 for Western blot). ** indicates statistically significant difference— p < 0.01.

Article Snippet: Ad-GFP (BD Biosciences) or Ad-PPARγ is intratracheally administrated into mice.

Techniques: Expressing, Isolation, Western Blot, Injection