Journal: Scientific Reports

Article Title: Monoacylglycerol Lipase: A Novel Potential Therapeutic Target and Prognostic Indicator for Hepatocellular Carcinoma

doi: 10.1038/srep35784

Figure Lengend Snippet: Methylation special primers were detected with MSP in all HCC cell lines but in L02 by MSP. Unmethylation special primers were only detected in L02. After treatment by DAC, methylation special primers decreased in HCC cell lines while unmethylation special primers increased ( A ). In HepG2 and SMMC-7721 cells ( B – D ), expression of LATS1 protein was suppressed. Demethylation of LATS1 promoter by DAC and knock-in LATS1 gene by plasmid transfection elevated LATS1 protein levels, which confirmed our findings that methylation of LATS1 promoter induced lost-expression of LATS1 protein in HCC. In HepG2 and SMMC-7721 cell lines, regulation of LATS1 with these methods induced significant changes in downstream signals YAP protein levels in a negatively correlative manner and pYAP protein levels in a positively correlative manner. Finally, due to methylation of LATS1 promoter and lost expression of LATS1, YAP protein could not be phosphorylated and detained in cytoplasm effectively. Excess YAP imported into nucleus and induced overexpression of MAGL.

Article Snippet: After transfer membranes were blocked with 5% nonfat milk, incubated with polyclonal antibodies for MAGL, LATS1, YAP1 or pYAP1 (Abcam, USA) respectively, and a monoclonal antibody for β-actin (Boster, China) was used as a loading control.

Techniques: Methylation, Expressing, Knock-In, Plasmid Preparation, Transfection, Over Expression

Journal: The Journal of Biological Chemistry

Article Title: Deletion of Monoglyceride Lipase in Astrocytes Attenuates Lipopolysaccharide-induced Neuroinflammation *

doi: 10.1074/jbc.M115.683615

Figure Lengend Snippet: MGL expression in MKO GFAP mice. A , Western blot analysis revealed absent MGL expression in lysates of primary cultivated astrocytes from MKO GFAP mice. Minor reductions were observed in lysates of MKO GFAP brains. MKO global brains served as control and showed no MGL expression. As controls we used floxed littermates for MKO GFAP mice and wild-type littermates for MKO global mice. B , MGL is expressed in primary neurons from cortex ( CTX ) and hippocampus ( HIP ) of MKO GFAP mice. Low expression was observed in primary microglia. Brain lysates of wild-type and MKO global mice served as positive and negative controls, respectively. C , immunofluorescence images of MGL ( red ) and GFAP ( green ) of brain cortical sections obtained from floxed control and MKO GFAP mice. Nuclei ( white ) were stained with DAPI. The scale bar represents 10 μm.

Article Snippet: Antibodies against MGL (polyclonal, in-house made rabbit anti-MGL serum , GFAP (ProSci, Flint Palace, CA), or GAPDH (Cell Signaling, Danvers, MA) and the respective horseradish peroxidase conjugated secondary antibodies were used.

Techniques: Expressing, Western Blot, Control, Immunofluorescence, Staining