Journal: Endocrinology

Article Title: Deficiency of FcϵR1 Increases Body Weight Gain but Improves Glucose Tolerance in Diet-Induced Obese Mice.

doi: 10.1210/en.2015-1184

Figure Lengend Snippet: Figure 4. Immunoblot analysis of adipogenesis-associated proteins in WAT from WT and Fcer1a/ mice after 17 weeks on a HFD. FcR1a immunoblot analysis (A), FcR1a immunostaining (B), and FcR1a immunofluorescent double staining together with Mac-2 or CD3 (C) of WAT from mice fed a chow diet and a HFD. Immunoblots or RT-PCR determined the expression of p-AKT and total AKT (D), C/EBP (E), PPAR (F), p- JNK and total JNK (G), Bcl-2 (H), and Glut4 (I) in WAT from WT and Fcer1a/ mice that consumed a HFD. -Actin immunoblots were used to ensure equal protein loading. Data are mean SEM of five independent experiments. Representative immunoblots for panels B-I are shown to the left.

Article Snippet: Antibody Table Protein Name Antibody Name Manufacturer, Catalog Number, or Name of Individual Provider Species Raised (Monoclonal or Polyclonal) Dilution Used Mac-2 Rabbit antimouse Mac 2 Cedarlane, 119–12296 Rabbit polyclonal 1:100 (IHC) CD3 Rabbit antihuman CD3 Abcam, ab16669 Rabbit monoclonal 1:60 (IHC) Fc R1 Antimouse Fc RI- eBioscience, 14-5898-82 Armenian Hamster monoclonal 1:50 (IHC) Fc R1 Antirabbit Fc RI- Upstate, 06–725 Rabbit polyclonal 1:1000 (IB) Bcl-2 Bcl-2 antibody Cell Signaling Technology, 2876S Rabbit polyclonal 1:1000 (IB) p-JNK Phospho-SAPK/JNK (Thr183/ Tyr185) (G9) mouse mAb Cell Signaling Technology, 5131 Mouse monoclonal 1:1000 (IB) JNK SAPK/JNK antibody Cell Signaling Technology, 9252S Rabbit polyclonal 1:500 (IB) p-AKT Phospho-Akt (Ser473) (D9E) XP rabbit mAb Cell Signaling Technology, 4060S Rabbit monoclonal 1:1000 (IB) AKT Akt antibody Cell Signaling Technology, 9272S Rabbit polyclonal 1:3000 (IB) Glut4 Glut4 antibody (C-20) Santa Cruz Biotechnology, sc-1608 Goat polyclonal 1:400 (IB) CEBP C/EBP antibody (14 AA) Santa Cruz Biotechnology, sc-61 Rabbit polyclonal 1:1000 (IB) PPAR PPAR antibody (H-100) Santa Cruz Biotechnology, sc-7196 Rabbit polyclonal 1:1000 (IB) UCP1 UCP1 antibody (C-17) Santa Cruz Biotechnology, sc-6528 Goat polyclonal 1:3000 (IB) -Actin Monoclonal anti- -actin Sigma, A5441 Mouse monoclonal 1:2000 (IB) Abbreviations: IB, immunoblot; IHC, immunohistochemistry; mAb, monoclonal antibody; SAPK, stress-activated protein kinase.

Techniques: Western Blot, Immunostaining, Double Staining, Reverse Transcription Polymerase Chain Reaction, Expressing

Journal: Endocrinology

Article Title: Deficiency of FcϵR1 Increases Body Weight Gain but Improves Glucose Tolerance in Diet-Induced Obese Mice.

doi: 10.1210/en.2015-1184

Figure Lengend Snippet: Figure 6. IgE activities in promoting adipocyte apoptosis and suppressing adipocyte glucose uptake. A, TUNEL staining and quantification of apoptotic cells in 3T3-L1 cells differentiated in the presence and absence of 50 g/mL IgE for indicated days. Representative data are shown to the right. B, Oil-red O staining of the same experiment from day 6 and day 8 experiments of panel A. C, TUNEL staining of preadipocytes and fully differentiated adipocyte treated with and without IgE (50 g/mL) to induce cell apoptosis. A CCK-8 assay determined cell viability (D) and an LDH assay determined cytotoxicity (E) of 3T3-L1 cells after 2 days of differentiation with and without different amount of IgE. F, Glucose uptake assay of 3T3-L1 cells after 2 days of differentiation with and without different amount of IgE. Preadipocytes were used as an experimental negative control. G, Glut4, p-AKT, and total AKT immunoblots of differentiated 3T3-L1 cells and treated with and without 50 g/mL IgE for 30 minutes. H, FcR1 siRNA- and scramble siRNA-transfected 3T3-L1 cells and differentiated for 4 days, followed by treatment with 25 g/mL of insulin or IgE for 10 minutes. -Actin immunoblots were used for protein loading controls. Data are mean SEM of three independent experiments.

Article Snippet: Antibody Table Protein Name Antibody Name Manufacturer, Catalog Number, or Name of Individual Provider Species Raised (Monoclonal or Polyclonal) Dilution Used Mac-2 Rabbit antimouse Mac 2 Cedarlane, 119–12296 Rabbit polyclonal 1:100 (IHC) CD3 Rabbit antihuman CD3 Abcam, ab16669 Rabbit monoclonal 1:60 (IHC) Fc R1 Antimouse Fc RI- eBioscience, 14-5898-82 Armenian Hamster monoclonal 1:50 (IHC) Fc R1 Antirabbit Fc RI- Upstate, 06–725 Rabbit polyclonal 1:1000 (IB) Bcl-2 Bcl-2 antibody Cell Signaling Technology, 2876S Rabbit polyclonal 1:1000 (IB) p-JNK Phospho-SAPK/JNK (Thr183/ Tyr185) (G9) mouse mAb Cell Signaling Technology, 5131 Mouse monoclonal 1:1000 (IB) JNK SAPK/JNK antibody Cell Signaling Technology, 9252S Rabbit polyclonal 1:500 (IB) p-AKT Phospho-Akt (Ser473) (D9E) XP rabbit mAb Cell Signaling Technology, 4060S Rabbit monoclonal 1:1000 (IB) AKT Akt antibody Cell Signaling Technology, 9272S Rabbit polyclonal 1:3000 (IB) Glut4 Glut4 antibody (C-20) Santa Cruz Biotechnology, sc-1608 Goat polyclonal 1:400 (IB) CEBP C/EBP antibody (14 AA) Santa Cruz Biotechnology, sc-61 Rabbit polyclonal 1:1000 (IB) PPAR PPAR antibody (H-100) Santa Cruz Biotechnology, sc-7196 Rabbit polyclonal 1:1000 (IB) UCP1 UCP1 antibody (C-17) Santa Cruz Biotechnology, sc-6528 Goat polyclonal 1:3000 (IB) -Actin Monoclonal anti- -actin Sigma, A5441 Mouse monoclonal 1:2000 (IB) Abbreviations: IB, immunoblot; IHC, immunohistochemistry; mAb, monoclonal antibody; SAPK, stress-activated protein kinase.

Techniques: TUNEL Assay, Staining, CCK-8 Assay, Lactate Dehydrogenase Assay, Negative Control, Western Blot, Transfection

Journal: Endocrinology

Article Title: Follistatin Targets Distinct Pathways To Promote Brown Adipocyte Characteristics in Brown and White Adipose Tissues

doi: 10.1210/en.2016-1607

Figure Lengend Snippet: List of Antibodies

Article Snippet: After the treatment with enhanced chemiluminescence reagent (Pierce; Thermo Scientific), the membranes were exposed to an X-ray film (Kodak; Sigma Aldrich, St. Louis, MO) and Image by Image Quant (GE Lifescience, Pittsburgh, PA) software for densitometric analysis ( 28, 32 ). table ft1 table-wrap mode="anchored" t5 Table 2. caption a7 Peptide/Protein Target Name of Antibody Manufacturer, Catalog Number Antibody RRID Species Dilution Used UCP1 Uncoupling protein1 antibody Abcam ab10983 AB_2241462 Rabbit polyclonal 1:1000 UCP2 Uncoupling protein2 antibody Novus Biologicals NB100-78377 AB_1085929 Rabbit polyclonal 1:1000 UCP3 Uncoupling protein3 antibody Abcam ab3477 AB_2304253 Rabbit polyclonal 1:1000 PRDM16 PRDM16 antibody (N16) Santa Cruz Biotech sc-130243 AB_2284344 Rabbit polyclonal 1:500 PGC-1 α PGC-1 α antibody (H-300) Santa Cruz Biotech sc-13067 AB_2166218 Rabbit polyclonal 1:1000 Adiponectin Adiponectin antibody R & D Systems MAB1119 AB_2305045 Rat monoclonal 1:1000 AMPK α AMPK α antibody Cell Signaling 2532 AB_330331 Rabbit monoclonal 1:1000 pAMPK α pAMPK α antibody Cell Signaling 2535 AB_331250 Rabbit monoclonal 1:1000 Glut4 Glucose transporter 4 antibody Abcam ab65267 AB_1140009 Mouse monoclonal 1:1000 p38MAPK p38MAPK antibody Cell Signaling 9212S AB_330713 Rabbit polyclonal 1:1000 pp38MAPK pp38MAPK antibody Cell Signaling 9211S AB_331641 Rabbit polyclonal 1:1000 ERK1/2 ERK1/2 antibody Cell Signaling 9102S AB_330744 Rabbit polyclonal 1:1000 pERK1/2 pERK1/2 antibody Cell Signaling 9101S AB_331646 Rabbit polyclonal 1:1000 Myf5 Myf5 antibody Santa Cruz Biotech sc-302 AB_631994 Rabbit polyclonal 1:1000 Cd137 Cd137 antibody Abcam ab3169 AB_303572 Mouse monoclonal 1:1000 BMP7 BMP7 antibody Abcam ab54904 AB_940598 Mouse monoclonal 1:1000 Eva1 Eva1 antibody Novus Biologicals NBP1-88937 AB_11016118 Rabbit polyclonal 1:1000 β -actin Beta actin antibody Santa Cruz Biotech sc-81178 AB_2223230 Mouse monoclonal 1:5000 GAPDH GAPDH antibody Millipore MAB374 AB_2107445 Mouse monoclonal 1:5000 pSmad2/3 pSmad2/3 antibody Cell Signaling 8828 AB_2631089 Rabbit monoclonal 1:1000 AcvRIIB AcvRIIB antibody Abcam ab76940 AB_1565807 Mouse monoclonal 1:1000 Smad3 Smad3 antibody Cell Signaling 9513 AB_2286450 Rabbit polyclonal 1:1000 Open in a separate window List of Antibodies Immunohistochemical analysis Embryo sections obtained from WT and Fst-Tg mice were fixed overnight in formalin, embedded with paraffin blocks and sectioned.

Techniques:

Journal: Endocrinology

Article Title: Follistatin Targets Distinct Pathways To Promote Brown Adipocyte Characteristics in Brown and White Adipose Tissues

doi: 10.1210/en.2016-1607

Figure Lengend Snippet: List of Antibodies

Article Snippet: After the treatment with enhanced chemiluminescence reagent (Pierce; Thermo Scientific), the membranes were exposed to an X-ray film (Kodak; Sigma Aldrich, St. Louis, MO) and Image by Image Quant (GE Lifescience, Pittsburgh, PA) software for densitometric analysis ( 28, 32 ). table ft1 table-wrap mode="anchored" t5 Table 2. caption a7 Peptide/Protein Target Name of Antibody Manufacturer, Catalog Number Antibody RRID Species Dilution Used UCP1 Uncoupling protein1 antibody Abcam ab10983 AB_2241462 Rabbit polyclonal 1:1000 UCP2 Uncoupling protein2 antibody Novus Biologicals NB100-78377 AB_1085929 Rabbit polyclonal 1:1000 UCP3 Uncoupling protein3 antibody Abcam ab3477 AB_2304253 Rabbit polyclonal 1:1000 PRDM16 PRDM16 antibody (N16) Santa Cruz Biotech sc-130243 AB_2284344 Rabbit polyclonal 1:500 PGC-1 α PGC-1 α antibody (H-300) Santa Cruz Biotech sc-13067 AB_2166218 Rabbit polyclonal 1:1000 Adiponectin Adiponectin antibody R & D Systems MAB1119 AB_2305045 Rat monoclonal 1:1000 AMPK α AMPK α antibody Cell Signaling 2532 AB_330331 Rabbit monoclonal 1:1000 pAMPK α pAMPK α antibody Cell Signaling 2535 AB_331250 Rabbit monoclonal 1:1000 Glut4 Glucose transporter 4 antibody Abcam ab65267 AB_1140009 Mouse monoclonal 1:1000 p38MAPK p38MAPK antibody Cell Signaling 9212S AB_330713 Rabbit polyclonal 1:1000 pp38MAPK pp38MAPK antibody Cell Signaling 9211S AB_331641 Rabbit polyclonal 1:1000 ERK1/2 ERK1/2 antibody Cell Signaling 9102S AB_330744 Rabbit polyclonal 1:1000 pERK1/2 pERK1/2 antibody Cell Signaling 9101S AB_331646 Rabbit polyclonal 1:1000 Myf5 Myf5 antibody Santa Cruz Biotech sc-302 AB_631994 Rabbit polyclonal 1:1000 Cd137 Cd137 antibody Abcam ab3169 AB_303572 Mouse monoclonal 1:1000 BMP7 BMP7 antibody Abcam ab54904 AB_940598 Mouse monoclonal 1:1000 Eva1 Eva1 antibody Novus Biologicals NBP1-88937 AB_11016118 Rabbit polyclonal 1:1000 β -actin Beta actin antibody Santa Cruz Biotech sc-81178 AB_2223230 Mouse monoclonal 1:5000 GAPDH GAPDH antibody Millipore MAB374 AB_2107445 Mouse monoclonal 1:5000 pSmad2/3 pSmad2/3 antibody Cell Signaling 8828 AB_2631089 Rabbit monoclonal 1:1000 AcvRIIB AcvRIIB antibody Abcam ab76940 AB_1565807 Mouse monoclonal 1:1000 Smad3 Smad3 antibody Cell Signaling 9513 AB_2286450 Rabbit polyclonal 1:1000 Open in a separate window List of Antibodies Immunohistochemical analysis Embryo sections obtained from WT and Fst-Tg mice were fixed overnight in formalin, embedded with paraffin blocks and sectioned.

Techniques: