Journal: Journal of Clinical Investigation

Article Title: GRP78 expression inhibits insulin and ER stress–induced SREBP-1c activation and reduces hepatic steatosis in mice

doi: 10.1172/jci37007

Figure Lengend Snippet: Figure 4 Expression of cytokines and ER stress markers in the livers of ob/+ and ob/ob mice injected with Ad GRP78 or Ad β-gal. Mice were injected with the indicated adenoviruses and sacrificed in the fed state 72 h later. (A) Total RNA was extracted and analyzed for IL-6 and TNF-α mRNA by qRT-PCR. *P < 0.05, **P < 0.01 versus respective ob/+ value; #P < 0.05 versus respective Ad β-gal value. (B) Microsomal membranes and nuclear extracts were prepared and analyzed by Western blot for the expression of precursor and nuclear ATF6, nuclear XBP-1, and microsomal GRP78. Each lane represents a different ani- mal. Results are representative of 3 independent experiments.

Article Snippet: The membranes were hybridized with the following antibodies: SREBP-1 (clone 2A4; Thermo Scientific), ATF6 (Imgenex), ChREBP (Novus Biological), XBP-1 (Prosci), and GRP78 and Insig-1 (Santa Cruz Biotechnology Inc.).

Techniques: Expressing, Injection, Quantitative RT-PCR, Western Blot

Journal: Journal of Experimental & Biomedical Sciences/Biomedical Science Letters

Article Title: Expression of Beta-catenin-interacting Protein 1 (CTNNBIP1) Gene Is Increased under Hypothermia but Decreased under Additional Ischemia Conditions

doi: 10.15616/bsl.2014.20.3.168

Figure Lengend Snippet: Fig. 3. Effect of hypothermia on gene expression of ER chaperones (A), XBP1 mRNA splicing (B), ER stress sensors (C), apoptosis- associated factors (D) and autophage-associated factors (E). PC12 cells were incubated under control conditions (37 ) or hypothermia ℃ (32 ) for 1 day. Gene expressions were determined by RT ℃ -PCR by conditions described in Materials and methods (A, D, E). The product of XBP1-Pst1 digestion was subjected to electrophoresis on the agarose gel (B). Both cleavage of ATF-6 and phosphorylation elF2α were assessed by Western blotting (C). The experiments were performed thrice and results represent the average and representative findings are shown. Bip; binding immunoglobulin protein, Canx; calnexin, PDI; protein disulfide isomerase, ATF6; activating transcription factor 6, phosph-elF2α; phosphorylation of eIF2α, Bak1; BCL2-antagonist/killer 1, Bcl-2; B-cell lymphoma 2, LC3; microtubule-associated protein 1 light chain 3α, beclin; coiled-coil myosin-like BCL2-interacting protein.

Article Snippet: Mouse anti-ATF6 antibody was obtained from Imgenex (San Diego, CA, USA).

Techniques: Gene Expression, Incubation, Control, Electrophoresis, Agarose Gel Electrophoresis, Phospho-proteomics, Western Blot, Binding Assay