Journal: The Journal of Biological Chemistry

Article Title: Physical and Functional Association of Lactate Dehydrogenase (LDH) with Skeletal Muscle Mitochondria *

doi: 10.1074/jbc.M113.476648

Figure Lengend Snippet: High-resolution confocal laser scanning microscopic imaging of immunolabeled LDH (red in A and B), IMM proteins (green in A) and Bcl-2 (green in B). Antibodies against Bcl-2 were used as a label of the outer mitochondrial membrane. Antibodies against the respiratory chain proteins were used to label the IMM. Pearson correlation coefficients for colocalization were 0.88 for IMM with LDH, and 0.73 for Bcl-2 with LDH. The scatter plots show the intensity of the red (Ch-1) and green (Ch-2) pixels.

Article Snippet: In intact fibers, scanning laser confocal microscopic imaging of antibodies specific for IMM proteins (mixture of antibodies against proteins of the respiratory chain, Mitosciences®) and LDH confirmed mitochondrial LDH colocalization ( A and A , a–c ).

Techniques: Imaging, Immunolabeling, Membrane

Journal: The Journal of Biological Chemistry

Article Title: Physical and Functional Association of Lactate Dehydrogenase (LDH) with Skeletal Muscle Mitochondria *

doi: 10.1074/jbc.M113.476648

Figure Lengend Snippet: Confocal laser scanning microscopic imaging of immunolabeled IMM and LDH of red gastrocnemius skeletal muscle fibers. Antibodies against proteins of the mitochondrial respiratory chain were used as a label for the IMM, along with antibodies against LDH. A, labeling of the intact fibers. B, labeled permeabilized fibers exhibiting attenuated LDH signal in the presence of the IMM signal. Note that the LDH signal was preserved in the SS mitochondria, which appear as larger elongated organelles at the cell edge. C, augmentation of the LDH signal by addition of the exogenous LDH (approximately 25 units/ml of LDH) + gentle wash. D, loss of the LDH staining in mitochondria with trypsin treatment.

Article Snippet: In intact fibers, scanning laser confocal microscopic imaging of antibodies specific for IMM proteins (mixture of antibodies against proteins of the respiratory chain, Mitosciences®) and LDH confirmed mitochondrial LDH colocalization ( A and A , a–c ).

Techniques: Imaging, Immunolabeling, Labeling, Gentle, Staining

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Phosphorylation is an on/off switch for 5-hydroxyconiferaldehyde O -methyltransferase activity in poplar monolignol biosynthesis

doi: 10.1073/pnas.1510473112

Figure Lengend Snippet: PtrAldOMT2 phosphopeptides identified by LC-MS/MS–based shotgun proteomic analysis of phosphopeptide-enriched P. trichocarpa SDX protein fractions. MS spectra: Ser123 (A) and Ser125 (B) phosphorylation. Underlined S represents phosphorylated serine residues. Fragment ions showing serine phosphorylation are labeled by dashed arrows. Numbers denote m/z values.

Article Snippet: The desalted phosphorylated SDX-recombinant protein mixtures were run on a Phosphate Affinity SDS/PAGE containing acrylamide-pendant Phos-tag (WAKO) using 100 V for 3 h at 4 °C.

Techniques: Liquid Chromatography with Mass Spectroscopy, Labeling

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Phosphorylation is an on/off switch for 5-hydroxyconiferaldehyde O -methyltransferase activity in poplar monolignol biosynthesis

doi: 10.1073/pnas.1510473112

Figure Lengend Snippet: PtrPAL1 and PtrPAL4|5 phosphopeptides identified by LC-MS/MS–based shotgun proteomic analysis of phosphopeptide enriched P. trichocarpa SDX protein fractions. MS spectra: PtrPAL1 (A) and PtrPAL4|5 (B) phosphorylation. Red S represents phosphorylated serine residues. Fragment ions showing serine phosphorylation are labeled by dashed arrows. Numbers next to peaks denote m/z values.

Article Snippet: The desalted phosphorylated SDX-recombinant protein mixtures were run on a Phosphate Affinity SDS/PAGE containing acrylamide-pendant Phos-tag (WAKO) using 100 V for 3 h at 4 °C.

Techniques: Liquid Chromatography with Mass Spectroscopy, Labeling

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Phosphorylation is an on/off switch for 5-hydroxyconiferaldehyde O -methyltransferase activity in poplar monolignol biosynthesis

doi: 10.1073/pnas.1510473112

Figure Lengend Snippet: (A) Purified recombinant PtrAldOMT2-6×His analyzed by SDS/PAGE (lane 1) stained with Coomassie-blue and BN-PAGE (lane 2) immunodetected by anti-His antibody. (B) P. trichocarpa SDX protein extracts separated by BN-PAGE and analyzed by in-gel activity assays using caffeic acid as substrate. (C) Confocal fluorescence imaging of P. trichocarpa SDX protoplasts expressing PtrAldOMT2–GFP or GFP-only control. (D) PtrAldOMT2 transcript abundance in fiber cells, vessel cells, and mixtures of three cell types (fiber, vessel, and ray cells). Error bars represent one SE of three technical replicates.

Article Snippet: The desalted phosphorylated SDX-recombinant protein mixtures were run on a Phosphate Affinity SDS/PAGE containing acrylamide-pendant Phos-tag (WAKO) using 100 V for 3 h at 4 °C.

Techniques: Purification, Recombinant, SDS Page, Staining, Activity Assay, Fluorescence, Imaging, Expressing

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Phosphorylation is an on/off switch for 5-hydroxyconiferaldehyde O -methyltransferase activity in poplar monolignol biosynthesis

doi: 10.1073/pnas.1510473112

Figure Lengend Snippet: (A) Recombinant PtrAldOMT2-6×His in vitro phosphorylation. (Lane 1) ATP is necessary for PtrAldOMT2 phosphorylation. (Lane 2) Ca2+ cannot facilitate PtrAldOMT2 phosphorylation. (Lane 3) Mg2+ weakly activates PtrAldOMT2 phosphorylation. (Lane 4) Mn2+ effectively activates PtrAldOMT2 phosphorylation. (Lane 5) CIAP dephosphorylates PtrAldOMT2. (B) P. trichocarpa SDX protein extracts activated recombinant PtrAldOMT2-6×His phosphorylation. Control assays, incubation at 0 °C; phosphorylation assays, incubation at 30 °C. Error bars represent one SE of five technical replicates. (C) Phos-tag SDS/PAGE of phospho-enrichment of recombinant PtrAldOMT2 phosphorylated by P. trichocarpa SDX, immunodetected by using anti-His antibody. (Lane 1) SDX-only control. (Lane 2) PtrAldOMT2-6×His only control. (Lane 3) Preenriched sample containing PtrAldOMT2-6×His and SDX. (Lane 4) Flow-through. (Lane 5) Wash buffer. (Lane 6) Enriched sample containing phosphorylated PtrAldOMT2-6×His and phosphorylated SDX proteins. (D) Activity of unphosphorylated and phosphorylated PtrAldOMT2-6×His, with caffeic acid (CA), caffealdehyde (CAFAld), or 5-hydroxyconiferaldehyde (5HCnAld) as substrate. Error bars represent one SE of three technical replicates.

Article Snippet: The desalted phosphorylated SDX-recombinant protein mixtures were run on a Phosphate Affinity SDS/PAGE containing acrylamide-pendant Phos-tag (WAKO) using 100 V for 3 h at 4 °C.

Techniques: Recombinant, In Vitro, Incubation, SDS Page, Activity Assay

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Phosphorylation is an on/off switch for 5-hydroxyconiferaldehyde O -methyltransferase activity in poplar monolignol biosynthesis

doi: 10.1073/pnas.1510473112

Figure Lengend Snippet: (A) ATP/Mn2+-activated phosphorylation of P. trichocarpa SDX protein extracts, analyzed by Phos-tag SDS/PAGE and immunodetected by using polyclonal anti-PtrAldOMT2 antibody. Control “C” denotes PtrAldOMT2-6×His. (B) AldOMT and PAL activities in phosphorylated, CIAP-treated, and untreated native P. trichocarpa SDX protein extracts. CA, caffeic acid; CAFAld, caffealdehyde; CiA, cinnamic acid; CnAld, coniferaldehyde; FA, ferulic acid; Phe, phenylalanine. Error bars represent one SE of three technical replicates. (C) Phos-tag SDS/PAGE of unphosphorylated recombinant PtrAldOMT2-6×His site-directed mutagenesis at Ser123 (S123N), Ser125 (S125N), and both Ser123 and Ser125 (S123N:S125N). (D) Phos-tag SDS/PAGE of in vitro phosphorylation of native and mutated PtrAldOMT2-6×His immunodetected by anti-His antibody. (Lane 1) PtrAldOMT2-6×His only control. (Lane 2) PtrAldOMT2-6×His in SDX. (Lane 3) S123N in SDX. (Lane 4) S125N in SDX. (Lane 5) S123N:S125N in SDX. (E) O-methyltransferase activity of native PtrAldOMT2-6×His, S123N, S125N, and S123N:S125N with caffeic acid, caffealdehyde, or 5-hydroxyconiferaldehyde as substrates. Activities are single measurement determinations.

Article Snippet: The desalted phosphorylated SDX-recombinant protein mixtures were run on a Phosphate Affinity SDS/PAGE containing acrylamide-pendant Phos-tag (WAKO) using 100 V for 3 h at 4 °C.

Techniques: SDS Page, Recombinant, Mutagenesis, In Vitro, Activity Assay

Journal: PLoS ONE

Article Title: Ildr1 gene deletion protects against diet-induced obesity and hyperglycemia

doi: 10.1371/journal.pone.0270329

Figure Lengend Snippet: (A) Ratio of caloric intake to body weight over 72 hr in mice fed the LFD. Light and dark cycles are represented as white or grey bars respectively over 24 hr intervals. (B) The results in panel A are expressed as the cumulative ratio for mice fed the LFD. (C) Caloric intake in mice given the LFD without normalization for body weight. (D) Cumulative time spent consuming the LFD diet [t(22) = 2.353, P = 0.028]. (E) Cumulative numbers of feeding bouts in LFD fed mice. (F) Ratio of caloric intake to body weight over 72 hr in the genotypes fed the HFD. (G) The results in panel F expressed as the cumulative ratio for mice fed the HFD [t(22) = 2.238, P = 0.036]. (H) Caloric intake in mice given the HFD without normalization for body weight [t(22) = 2.180, P = 0.040]. (I) Cumulative time spent consuming the HFD diet [t(22) = 2.417, P = 0.024]. (J) Cumulative numbers of feeding bouts in HFD groups [t(22) = 2.097, P = 0.048]. n = 12 mice/genotype/diet; *P<0.05, WT vs . Ildr1 -/- .

Article Snippet: The protein was dialyzed and concentrated, and a mixture of mouse and human ILDR1 proteins was used to immunize rabbits (PrimmBiotech Inc., Cambridge MA).

Techniques:

Journal: PLoS ONE

Article Title: Ildr1 gene deletion protects against diet-induced obesity and hyperglycemia

doi: 10.1371/journal.pone.0270329

Figure Lengend Snippet: Mice were fed the low fat or the high fat diet for 18 days after removal from the CLAMS apparatus (total of 23 days). (A) Body weight of mice fed the LFD. (B) Body weight of mice fed the HFD [t(6) = 3.697, P = 0.010]. n = 4 mice/genotype/diet; *P<0.05, WT vs. Ildr1 -/- .

Article Snippet: The protein was dialyzed and concentrated, and a mixture of mouse and human ILDR1 proteins was used to immunize rabbits (PrimmBiotech Inc., Cambridge MA).

Techniques:

Journal: PLoS ONE

Article Title: Ildr1 gene deletion protects against diet-induced obesity and hyperglycemia

doi: 10.1371/journal.pone.0270329

Figure Lengend Snippet: (A) Motor activities over 72 hr with 12-hr light and dark cycles (cycles represented as white or grey bars respectively) in wild-type and Ildr1 -/- mice fed the LFD. (B) Cumulative beam-breaks over 72 hr in the LFD groups [t(22) = 2.127, P = 0.045]. (C) Energy expenditure (heat production) over 72 hr with 12-hr light and dark cycles in the wild-type and Ildr1 -/- mice fed the LFD. (D) Cumulative energy expenditure over 72 hr in the LFD groups. (E) Motor activities over 72 hr in the wild-type and Ildr1 -/- mice fed the HFD. (F) Cumulative beam-breaks over 72 hr in the HFD groups [t(22) = 3.097, P = 0.005]. (G) Energy expenditure over 72 hr with 12-hr light and dark cycles in the HFD groups. (H) Cumulative energy expenditure over 72 hr in the HFD groups [t(22) = 2.744, P = 0.012]. n = 12 mice in each group; *P<0.05, **P<0.01, WT vs . Ildr1 -/- .

Article Snippet: The protein was dialyzed and concentrated, and a mixture of mouse and human ILDR1 proteins was used to immunize rabbits (PrimmBiotech Inc., Cambridge MA).

Techniques:

Journal: PLoS ONE

Article Title: Ildr1 gene deletion protects against diet-induced obesity and hyperglycemia

doi: 10.1371/journal.pone.0270329

Figure Lengend Snippet: (A) VO 2 intake over 72 hr with 12-hr light and dark cycles represented as gray or black bars, respectively, in the LFD groups. (B) Cumulative VO 2 intake during the dark cycle in the LFD groups. (C) VCO 2 exhalation over 72 hr with 12-hr light and dark cycles in the LFD groups. (D) Cumulative VCO 2 output during the dark cycle in the LFD groups. (E) VO 2 intake across circadian cycles in the HFD groups. (F) Cumulative VO 2 intake during the dark cycle in the HFD groups [t(22) = 2.402, P = 0.025]. (G) VCO 2 exhalation over 72 hr with12 hr light and dark cycles. (H) Cumulative VCO 2 output during the dark cycle in the HFD groups [t(22) = 2.324, P = 0.030]. n = 12 mice/genotype/diet; *P<0.05, WT vs . Ildr1 -/- .

Article Snippet: The protein was dialyzed and concentrated, and a mixture of mouse and human ILDR1 proteins was used to immunize rabbits (PrimmBiotech Inc., Cambridge MA).

Techniques:

Journal: PLoS ONE

Article Title: Ildr1 gene deletion protects against diet-induced obesity and hyperglycemia

doi: 10.1371/journal.pone.0270329

Figure Lengend Snippet: Four adipose depots were collected in mice fed either a low fat (LFD) or high fat diet (HFD). (A, E) Epididymal adipose tissue weight to body weight in LFD (A) or HFD (E) fed mice. (B, F) Inguinal adipose tissue weight to body weight in LFD (B) or HFD (F) fed mice. (C, G) Retroperitoneal adipose tissue weight to body weight in LFD (C) or HFD (G) fed mice. (D, H) Brown adipose tissue weight to body weight in LFD (D) or HFD (H) fed mice; for mice fed the HFD group [t(6) = 3.590, P = 0.012]. n = 3–4 mice/genotype/diet; *P<0.05, WT vs . Ildr1 -/- .

Article Snippet: The protein was dialyzed and concentrated, and a mixture of mouse and human ILDR1 proteins was used to immunize rabbits (PrimmBiotech Inc., Cambridge MA).

Techniques:

Journal: PLoS ONE

Article Title: Ildr1 gene deletion protects against diet-induced obesity and hyperglycemia

doi: 10.1371/journal.pone.0270329

Figure Lengend Snippet: (A) LFD fed mice: 1001–2000 μm 2 cells [t(6) = 5.373, P = 0.0017]; 3001–4000 μm 2 cells [t(6) = 4.259, P = 0.0054], 4001–5000 μm 2 cells [t(6) = 7.999, P = 0.0002], 5001–6000 μm 2 cells [t(6) = 7.462, P = 0.0003], 6001–7000 μm 2 cells [t(6) = 3.892, P = 0.0081]; (B) HFD fed mice: <1000 μm 2 cells [t(5) = 2.929, P = 0.032], 1001–2000 μm 2 [t(5) = 6.22, P = 0.0015], 2001–3000 μm 2 cells [t(5) = 3.811, P = 0.012], 6001–7000 μm 2 cells [t(5) = 2.804, P = 0.037]. n = 3–4 mice/genotype/diet; *P<0.05, **P<0.01, ***P<0.001, WT vs . Ildr1 -/- .

Article Snippet: The protein was dialyzed and concentrated, and a mixture of mouse and human ILDR1 proteins was used to immunize rabbits (PrimmBiotech Inc., Cambridge MA).

Techniques:

Journal: PLoS ONE

Article Title: Ildr1 gene deletion protects against diet-induced obesity and hyperglycemia

doi: 10.1371/journal.pone.0270329

Figure Lengend Snippet: (A, D) Adiponectin levels in LFD or HFD fed mice; for LFD group [t(8) = 4.130, P = 0.003]. (B, E) Leptin levels in LFD or HFD fed mice; for the LFD group [t(8) = 7.496, P<0.0001]. (C, F) T4 hormone levels in LFD or HFD fed mice. n = 4–6 mice/genotype or diet; **P<0.01, ****P<0.0001, WT vs . Ildr1 -/- .

Article Snippet: The protein was dialyzed and concentrated, and a mixture of mouse and human ILDR1 proteins was used to immunize rabbits (PrimmBiotech Inc., Cambridge MA).

Techniques:

Journal: PLoS ONE

Article Title: Ildr1 gene deletion protects against diet-induced obesity and hyperglycemia

doi: 10.1371/journal.pone.0270329

Figure Lengend Snippet: (A-C) Glucagon (green) and insulin (red) immunostaining in wild-type islets. (A) Glucagon positive alpha cells (green) are present primarily in the periphery of the islet. (B) Insulin positive beta cells (red) comprise the majority of the islet. (C) Merged image of glucagon and insulin immunostaining in a wild-type islet. (D-F) Glucagon (red) and β-galactosidase reporter (GLB1) (green) immunostaining in wild-type islet. (D) A few glucagon expressing cells (red) are present in the periphery of wild-type islets. (E) This islet is negative for β-galactosidase reporterimmunostaining. (F) Merged image of glucagon and β-galactosidase reporter immunostaining in wild-type islet. (G-I) Glucagon (red) and β-galactosidase reporter (green) immunostaining in Ildr1 -/- islet. (G) Similar to wild-type islet, glucagon expressing cells (red) are present in the periphery of Ildr1 -/- islet. (H) However, unlike wild-type islet, in Ildr1 -/- mice, β-galactosidase reporter (green) expression is detected. (I) Glucagon and β-galactosidase reporterantigens appear to co-localize (yellow) in the merged image. β-galactosidase reporter expression was not observed in acinar cells that surround the islets, in accordance with X-gal staining (shown in above). Scale bar is 30 μm in all the panels.

Article Snippet: The protein was dialyzed and concentrated, and a mixture of mouse and human ILDR1 proteins was used to immunize rabbits (PrimmBiotech Inc., Cambridge MA).

Techniques: Immunostaining, Expressing, Staining

Journal: PLoS ONE

Article Title: Ildr1 gene deletion protects against diet-induced obesity and hyperglycemia

doi: 10.1371/journal.pone.0270329

Figure Lengend Snippet: (A-C) Insulin (red) and β-galactosidase reporter (green) staining in wild-type (WT) islets. (A) Insulin (red) expressing beta cells are present in the WT islet. (B) These cells are negative for the β-galactosidase reporter (green). (C) Merged image of insulin and β-galactosidase reporter immunostaining. (D-F) Insulin (red) and β-galactosidase reporter (green) staining in Ildr1 -/- islet. (D) Insulin (red) expressing beta cells are also present in the Ildr1 -/- islet. (E) β-galactosidase reporter (green) is expressed in Ildr1 -/- islet. (F) Merged image of insulin and β-galactosidase reporter immunostaining showing colocalization (yellow). (G-I) Insulin (green) and ILDR1 (red) immunostaining in wild-type (WT) islet. (G) Insulin immunostaining (green) in WT islet. (H) ILDR immunostaining (red) in WT islet. (I) While the ILDR1 immunostaining is faint, it co-localizes with insulin in islets of WT mice. (J-L) A high resolution airy scan image of insulin (green) and ILDR1 (red) in islet of WT mice. (J) Insulin immunostaining at higher magnification and resolution. (K) Corresponding ILDR1 immunostaining (green) (L) Insulin and ILDR1 appear to be spatially segregated within beta cells. (M-O) Insulin (green) and ILDR1 (red) immunostaining in Ildr1 -/- islet. (M) Insulin-expressing beta cells (green) are abundant in Ildr1 -/- islet. (N) ILDR1 immunostaining is absent in Ildr1 -/- islet. (O) Merged image of insulin and ILDR1 immunostaining. Scale bars are 30 μm for images showing whole islet and 10 μm for the high resolution images (panels J-L).

Article Snippet: The protein was dialyzed and concentrated, and a mixture of mouse and human ILDR1 proteins was used to immunize rabbits (PrimmBiotech Inc., Cambridge MA).

Techniques: Staining, Expressing, Immunostaining

Journal: PLoS ONE

Article Title: Ildr1 gene deletion protects against diet-induced obesity and hyperglycemia

doi: 10.1371/journal.pone.0270329

Figure Lengend Snippet: All tests were performed under low fat diet condition. (A) Oral glucose tolerance test (OGTT) in wild-type (WT, grey lines and grey bars) and Ildr1 -/- mice (black lines and black bars); RMANOVA for time [F(5,40) = 45.183, P<0.001], genotype [F(1,8) = 8.115, P = 0.022], and time by genotype [F(5,40) = 2.881, P = 0.026]. Inset : Area under the curve (AUC) for OGTT [t(8) = 3.541, P = 0.008]; n = 5 mice/genotype. (B) Glucose excursion in the mixed meal tolerance test (MMTT) in WT and Ildr1 -/- mice: RMANOVA for time [F(6,60) = 11.601, P<0.001], genotype [F(1,10) = 7.112, P = 0.024], and time by genotype [F(6,60) = 2.426, P = 0.036]; Inset : AUC for MMTT [t(10) = 2.335, P = 0.042]; n = 6 mice/genotype. (C) MMTT insulin measured at 0 and 10 min: step-wise regression for time [R 2 = 0.048; F(1,23) = 1.120, P = 0.301] and genotype from baseline (time 0) to 10 min [R 2 = 0.320; F(2,23) = 4.952, P = 0.017]. (D) Glucose excursion in the insulin tolerance test (ITT) in WT (grey lines/grey bars) and Ildr1 -/- mice (black lines/black bars). n = 6 mice/genotype. (E) Insulin tolerance test (ITT) monitoring insulin secretion from ex vivo glucose- or KCl-perfused WT and Ildr1 -/- islets (n = 6 mice/genotype). (F) Integrated area under the curve for glucose-stimulated insulin secretion ex vivo (8–24 min; from panel E) and KCl-stimulated insulin secretion (40–56 min); for glucose [t(22) = 3.082, P = 0.005, two-tailed]. *P<0.05, **P<0.01 WT vs . Ildr1 -/- .

Article Snippet: The protein was dialyzed and concentrated, and a mixture of mouse and human ILDR1 proteins was used to immunize rabbits (PrimmBiotech Inc., Cambridge MA).

Techniques: Ex Vivo, Two Tailed Test

Journal: PLoS ONE

Article Title: Ildr1 gene deletion protects against diet-induced obesity and hyperglycemia

doi: 10.1371/journal.pone.0270329

Figure Lengend Snippet: Metabolite profiles of wild-type (WT) and Idlr1 -/- mice fed a low fat diet (LFD) and high fat diet (A) Adipose amino acid profile for Ala: diet [F(1,22) = 4.436, P = 0.047]; for Asx: diet [F(1,22) = 14.686, P<0.001], genotype by diet [F(1,22) = 6.057, P = 0.022]; and for Glx: diet [F(1,22) = 13.319, P = 0.001], genotype by diet [F(1,22) = 4.295, P = 0.050]; for Pro: diet [F(1,22) = 12.128, P = 0.002], genotype by diet [F(1,22) = 5.753, P = 0.025]; for Val: genotype by diet [F(1,22) = 4.971, P = 0.036]; Ile/Leu: genotype by diet [F(1,22) = 6.604, P = 0.017]; for His: genotype by diet [F(1,22) = 5.645, P = 0.027]; Phe: genotype by diet [F(1,22) = 4.495, P = 0.045]; and for Tyr: diet [F(1,22) = 9.309, P = 0.006], genotype by diet [F(1,22) = 4.351, P = 0.049]. (B) Blood spot amino acid profile for Tyr: genotype [F(1,23) = 7.552, P = 0.011], diet [F(1,23) = 17.184, P<0.001], genotype by diet [F(1,23) = 11.497, P = 0.003]. (C) Liver amino acid profile for Gly: diet [F(1,23) = 35.189, P<0.001]; for Met: diet [F(1,23) = 16.715, P<0.001]; for Tyr: diet [F(1,23) = 4.528, P = 0.044]; for Orn: genotype [F(1,23) = 4.192, P = 0.052], diet [F(1,23) = 16.240, P<0.001]; for Cit: genotype [F(1,23) = 10.913, P = 0.003], diet [F(1,23) = 11.373, P = 0.003]; and for Arg: genotype [F(1,23) = 7.633, P = 0.011], diet [F(1,23) = 10.412, P = 0.004]. (D) Liver organic acid profile for αKg: genotype by diet [F(1,21) = 4.876, P = 0.038]; for Pyr: diet [F(1,23) = 5.571, P = 0.027], and for Cit: diet [F(1,23) = 5.857, P = 0.024], genotype by diet [F(1,23) = 4.227, P = 0.051]. n = 5–8 samples/group. *P<0.05, **P<0.01, ***P<0.001, WT vs . Ildr1 -/- within diet; +P<0.05, ++P<0.01, +++P<0.001, low vs . high fat diet within genotype; ^P<0.05, ^^P<0.01, overall genotype effect; #P<0.05, ##0.01, ###P<0.001, overall diet effect.

Article Snippet: The protein was dialyzed and concentrated, and a mixture of mouse and human ILDR1 proteins was used to immunize rabbits (PrimmBiotech Inc., Cambridge MA).

Techniques: