anti-o1 antibody  (Thermo Fisher)

Journal: Nutrients

Article Title: EGCG and Taurine Synergistically Ameliorate Lipid Metabolism Disorder by Modulating Gut Microbiota and PPARα/FAS Signaling Pathway

doi: 10.3390/nu17162595

Figure Lengend Snippet: Effects of EGCG and taurine on the expression of proteins related to fatty acid synthesis and catabolism. ( A ) Fox-o1 and PPARα protein bands; ( B ) FAS and LPL protein bands. ( C ) Relative expression level of PPARα; ( D ) relative expression level of FAS; ( E ) relative expression level of Fox-o1; ( F ) relative expression level of LPL; NC: normal diet control, MC: high-fat emulsion established model control, PC: simvastatin treatment positive control, EC: EGCG treatment, TT: taurine treatment, ETC: combined EGCG and taurine treatment. Data are means ± SEM (n = 2). a–e: Different letters in the same column indicate significant differences in the numerical values ( p < 0.05).

Article Snippet: Antibodies against fatty acid synthase (FAS), peroxisome proliferator-activated receptor alpha (PPARα), forkhead box protein o1 (Fox-o1), and lipoprotein lipase (LPL) were purchased from Proteintech Biotechnology Co., Ltd. (Wuhan, China).

Techniques: Expressing, Control, Emulsion, Positive Control

Journal: World Journal of Gastroenterology

Article Title: Kushenol I combats ulcerative colitis via intestinal barrier preservation and gut microbiota optimization

doi: 10.3748/wjg.v31.i26.105656

Figure Lengend Snippet: Molecular docking and corresponding scores of kushenol I with key targets protein kinase B, p38 mitogen-activated protein kinase, NOD-like receptor thermal protein domain associated protein 3, phosphoinositide 3-kinase, protein kinase B, forkhead box O1, and Toll-like receptor 4, are presented. AKT: Protein kinase B; FOXO1: Forkhead box O1; NF-κB: Nuclear factor kappa B; NLRP3: NOD-like receptor thermal protein domain associated protein 3; p38 MAPK: p38 mitogen-activated protein kinase; PI3K: Phosphoinositide 3-kinase; TLR4: Toll-like receptor 4.

Article Snippet: Antibodies targeting forkhead box O1 (FOXO1), p38 mitogen-activated protein kinase (p38 MAPK), phosphorylated p38 MAPK (p-p38 MAPK), protein kinase B (AKT), phosphorylated protein kinase B (p-AKT), and β-actin were sourced from Proteintech (Proteintech, Rosemont, IL, United States).

Techniques:

Journal: World Journal of Gastroenterology

Article Title: Kushenol I combats ulcerative colitis via intestinal barrier preservation and gut microbiota optimization

doi: 10.3748/wjg.v31.i26.105656

Figure Lengend Snippet: Molecular dynamics simulation results of kushenol I interacting with protein kinase B, forkhead box O1, nuclear factor kappa B, NOD-like receptor thermal protein domain associated protein 3, phosphoinositide 3-kinase, p38 mitogen-activated protein kinase, and Toll-like receptor 4. A: Fluctuation curves of hydrogen bond numbers; B: Radius of gyration curves; C: Root mean square deviation curves; D: Root mean square fluctuation curves; E: Free energy landscape. AKT: Protein kinase B; FOXO1: Forkhead box O1; NF-κB: Nuclear factor kappa B; NLRP3: NOD-like receptor thermal protein domain associated protein 3; p38 MAPK: p38 mitogen-activated protein kinase; PI3K: Phosphoinositide 3-kinase; Rg: Gyration radius; RMSD: Root mean square deviation; RMSF: Root mean square deviation; TLR4: Toll-like receptor 4.

Article Snippet: Antibodies targeting forkhead box O1 (FOXO1), p38 mitogen-activated protein kinase (p38 MAPK), phosphorylated p38 MAPK (p-p38 MAPK), protein kinase B (AKT), phosphorylated protein kinase B (p-AKT), and β-actin were sourced from Proteintech (Proteintech, Rosemont, IL, United States).

Techniques:

Journal: World Journal of Gastroenterology

Article Title: Kushenol I combats ulcerative colitis via intestinal barrier preservation and gut microbiota optimization

doi: 10.3748/wjg.v31.i26.105656

Figure Lengend Snippet: Effects of kushenol I on the expression of inflammation-related proteins phosphorylated phosphoinositide 3-kinase (PI3K), PI3K, phosphorylated protein kinase B (AKT), AKT, forkhead box O1, interleukin 1β, Toll-like receptor 4, phosphorylated p38 mitogen-activated protein kinase (MAPK), p38 MAPK, nuclear factor kappa B phosphorylated p65, nuclear factor kappa B p65, and NOD-like receptor thermal protein domain associated protein 3 in the colonic tissues of mice with ulcerative colitis. A: Image of a gel showing protein expression; B: Bar graph depicting protein expression. Values are presented as the mean ± standard error of the mean ( n = 3). a P < 0.05. b P < 0.01. c P < 0.001. P compared with the model group. AKT: Protein kinase B; FOXO1: Forkhead box O1; IL-1β: Interleukin 1β; PI3K: Phosphoinositide 3-kinase; p-p38: Phosphorylated p38; p38 MAPK: p38 mitogen-activated protein kinase; p-p65: Phosphorylated p65; p-PI3K: Phosphorylated phosphoinositide 3-kinase; p-AKT: Phosphorylated protein kinase B; NF-κB: Nuclear factor kappa B; NLRP3: NOD-like receptor thermal protein domain associated protein 3; TLR4: Toll-like receptor 4.

Article Snippet: Antibodies targeting forkhead box O1 (FOXO1), p38 mitogen-activated protein kinase (p38 MAPK), phosphorylated p38 MAPK (p-p38 MAPK), protein kinase B (AKT), phosphorylated protein kinase B (p-AKT), and β-actin were sourced from Proteintech (Proteintech, Rosemont, IL, United States).

Techniques: Expressing

Journal: World Journal of Gastroenterology

Article Title: Kushenol I combats ulcerative colitis via intestinal barrier preservation and gut microbiota optimization

doi: 10.3748/wjg.v31.i26.105656

Figure Lengend Snippet: Kushenol I preserves the intestinal barrier and optimizes gut microbiota to combat ulcerative colitis. AKT: Protein kinase B; CD: Cluster of differentiation; FOXO1: Forkhead box O1; GSH-PX: Glutathione peroxidase; IL: Interleukin; MDA: Malondialdehyde; MPO: Myeloperoxidase; NF-κB: Nuclear factor kappa B; NLRP3: NOD-like receptor thermal protein domain associated protein 3; PI3K: Phosphoinositide 3-kinase; SOD: Superoxide dismutase; TNF: Tumor necrosis factor; TLR4: Toll-like receptor 4; ZO-1: Zonula occludens-1.

Article Snippet: Antibodies targeting forkhead box O1 (FOXO1), p38 mitogen-activated protein kinase (p38 MAPK), phosphorylated p38 MAPK (p-p38 MAPK), protein kinase B (AKT), phosphorylated protein kinase B (p-AKT), and β-actin were sourced from Proteintech (Proteintech, Rosemont, IL, United States).

Techniques:

Journal: Journal of Neurochemistry

Article Title: Gα o1 and Gα o1 /Gα o2 deletion differentially affect hippocampal mossy fiber tract anatomy and neuronal morphogenesis

doi: 10.1111/jnc.16248

Figure Lengend Snippet: Altered projections and reduced size of mossy fiber synaptic areas in the hippocampus of Gα o1 −/− mice. (a) Coronal section from an adult wild‐type mouse brain was immunostained for the vesicular Zinc transporter 3 (ZnT3, heavily marking the mossy fiber tract (mft), and the axonal marker neurofilament heavy chain (NFP)), together with Dapi to depict the gross anatomy of the hippocampus and its layered organization. The mossy fiber tract originates from the granule cells of the dentate gyrus, passes through the hilus, and terminates at the Stratum lucidum (SLU) forming synapses with the CA3 pyramidal cells. (b) Schematic representation of the hippocampus formation and the mossy fiber tract bundles. Before reaching the Stratum lucidum , the mossy fiber tract splits up into two bundles running above and below the pyramidal cells. Arrows indicate the two distance lines measured to determine the gross size of the hippocampus formation in the medio‐lateral (width) and dorso‐ventral (height) axis. GCL, granule cell layer; HIL, hilum ; SLM, Stratum lacunosum/moleculare ; SM, Stratum moleculare; SO, Stratum oriens ; SP, Stratum pyramidale ; SR, Stratum radiatum ; (c) Representative section of a wild‐type hippocampus stained for ZnT3 to demonstrate the mossy fiber tract substructures analyzed for quantification. Mossy fibers project from the hilus above and below the pyramidal cell layer, termed suprapyramidal and infrapyramidal bundle (SPB and IPB) before terminating in the Stratum lucidum (SLU). Length of the IPB, SPB, SPB + SLU, SLU alone as well as the overall length of the tract from the tip of the hilus to the tip of the SLU were determined (see yellow lines for portions of the tract). Additionally, the area of the SLU and the rest of the mossy fiber tract were quantified. (d, e) Representative images of adult wild‐type (same as in c) and a Gα o1 −/− hippocampi stained for ZnT3 are shown. Most strikingly, the MFT appears thinner and the IPB appeared to be longer in the knockout animals than in the wild‐type. (f, g) Quantification of measurements. Both the SLU and overall synaptic mossy fiber area were markedly reduced in Gα o1 −/− animals. General hippocampus width was not significantly altered, hippocampus height was slightly reduced in KO animals. Most prominently, the length of the IPB was reduced in wild‐type compared to the knockout animals, accompanied by a slight reduction in the SBP length. The length of the SLU was moderately reduced in G α O1 knockout animals. The other parameters were unaltered. Bars show means ± SD from N = 4 pooled animals each WT and Gα o1 −/− , 51 (knockout) and 54 (wild‐type) mossy fiber tracts per genotype *** p ≤ 0.001 (h) Confocal imaging and 3D reconstruction from z‐stacks showing ZnT3 expression together with surrounding axons stained by NFP at a wild‐type SLU area (left panel). 3D reconstruction of ZnT3 staining at the SLU area showing the differences in SLU size and IPB length between wild‐type (middle panel) and Gα o1 −/− mice (right panel).

Article Snippet: A polyclonal antibody preferentially recognizing Gα o1 was from Santa Cruz (cat. no. 13532, Santa Cruz, CA, USA).

Techniques: Marker, Staining, Knock-Out, Imaging, Expressing

Journal: Journal of Neurochemistry

Article Title: Gα o1 and Gα o1 /Gα o2 deletion differentially affect hippocampal mossy fiber tract anatomy and neuronal morphogenesis

doi: 10.1111/jnc.16248

Figure Lengend Snippet: Knockout of Gα o1 as verified by genotyping, western blot, and immunohistochemistry. (a) Exemplary genotyping of wild‐type and Gα o1 heterozygous and homozygous knockout animals. Genomic template DNA was isolated from ear punches and amplified by PCR. In the wild‐type and heterozygous mice, Gα o1 DNA was clearly detectable but absent in homozygous mice. Neomycin (Neo) cassette DNA confirmed knockout. (b) Western blot analysis of Gα o1 and Gα o2 protein expression. Brain homogenates of wild‐type and Gα o1 −/− mice were stained by an antibody preferentially recognizing Gα o1 and an antibody detecting both Gα o1 and Gα o2 . Vesicular Synaptobrevin (Syb) was used as control protein. Both Gα o antibodies detected a major band below 40 kDa in the wild‐type that was largely absent in the knockout when using the antibody preferentially detecting Gα o1 and still detected by the Gα o1/2 antibody. (c) Immunohistochemical analysis of Gα o1 and Gα o2 expression in wild‐type and Gα o1 −/− mice. Coronal brain sections of adult mice of either strain were stained with the same antibodies as shown above. In the wild‐type, both antibodies showed a ubiquitous immune reaction in virtually all brain areas. Again, immunostaining against Gα o1 was largely absent in the knockout, while Gα o2 was still clearly detectable by the Gα o1/2 antibody (see insets for hippocampal CA3 area).

Article Snippet: A polyclonal antibody preferentially recognizing Gα o1 was from Santa Cruz (cat. no. 13532, Santa Cruz, CA, USA).

Techniques: Knock-Out, Western Blot, Immunohistochemistry, Isolation, Amplification, Expressing, Staining, Control, Immunohistochemical staining, Immunostaining

Journal: Journal of Neurochemistry

Article Title: Gα o1 and Gα o1 /Gα o2 deletion differentially affect hippocampal mossy fiber tract anatomy and neuronal morphogenesis

doi: 10.1111/jnc.16248

Figure Lengend Snippet: Development of differences in mossy fiber tract anatomy between wild‐type and Gα o1 −/− mice. Measurements of mossy fiber tract parameters of wild‐type and Gα o1 −/− mice from postnatal day 2 into adulthood. Analysis was based on either ZnT3 (P12, P16) or Synaptoporin (P2‐P8) stainings. (a) At P2, only very slight differences in the size of the synaptic area of the SLU and the total length of the mossy fiber tract were observed. All other parameters analyzed remained unchanged. (b) At P16, changes of analyzed parameters in Gα o1 −/− mice were most pronounced for reduction in synaptic areas and increased length of IBP. Bars show means ± SD from N = 3 pooled animals each WT and G α o1 −/− per age, 30 mossy fiber tracts per genotype and age. * p ≤ 0.05; *** p ≤ 0.001 (c–h) Absolute changes displayed in (a) and (b) and Figure for the individual time points are summarized as relative changes from P2 to adulthood. Values for P2 (both for wild‐type and Gα o1 −/− mice) were set as 1. (c) Fold changes in SLU area. (d) Fold changes in total MFT area. (e) Fold changes in IPB length. (f) Fold changes in total MFT length. (g) Fold changes in SLU length. (h) Fold changes in hippocampus dimensions. Data show means ± SD, N values correspond to the information given in a and b.

Article Snippet: A polyclonal antibody preferentially recognizing Gα o1 was from Santa Cruz (cat. no. 13532, Santa Cruz, CA, USA).

Techniques:

Journal: Journal of Neurochemistry

Article Title: Gα o1 and Gα o1 /Gα o2 deletion differentially affect hippocampal mossy fiber tract anatomy and neuronal morphogenesis

doi: 10.1111/jnc.16248

Figure Lengend Snippet: Double knockout of Gα o1 and Gα o2 largely restores inhibitory effects of single Gα o1 deletion on synaptic area and alterations in mossy fiber tract substructure length. (a) Exemplary genotyping of wild‐type and Gα o (Gα o1 and Gα o2 ) heterozygous and homozygous knockout animals. Genomic template DNA was isolated from ear punches and amplified by PCR. In the wild‐type and heterozygous mice, Gα o DNA was clearly detectable but absent in homozygous mice. Presence of Neomycin (Neo) cassette DNA confirms knockout in heterozygous and homozygous animals. (b) Western blot analysis of Gα o1 and Gα o2 protein expression. Brain homogenates of wild‐type and Gα o −/− mice were stained by an antibody preferentially recognizing Gα o1 and an antibody detecting both Gα o1 and Gα o2 . Vesicular Synaptobrevin (Syb) was used as internal control protein. Both Gα o antibodies detected a major band below 40 kDa in the wild‐type that was fully absent in the knockout when using either antibody. (c) Immunohistochemical analysis of Gα o1 and Gα o2 expression in wildtype and Gα o −/− mice. Coronal brain sections of adult mice of either strain were stained with the same antibodies as shown above. In the wild‐type, both Gα o1 and Gα o1/2 antibodies showed a ubiquitous signal in virtually all brain areas. Again, immunostaining against Gα o1 and Gα o2 was negative for both antibodies in the knockout. (d, e) Quantification of mossy fiber tract measurements of adult and P2 wild‐type and Gα o −/− mice based on Synaptoporin stainings. In the adult, total MFT synaptic area and, very moderately, hippocampus height were reduced in knockout animals. All other parameters remained unchanged in the knockout. At P2, moderate changes appeared in the synaptic SLU area and all MFT length parameters with slightly higher values in the knock‐out. Bars show means ± SD from N = 4/3 pooled animals each WT and Gα o −/− adult/P2, 40/30 mossy fiber tracts per adult/P2 genotype * p ≤ 0.05; ** p ≤ 0.01; *** p ≤ 0.001 (f) Representative coronal sections of adult wild‐type and Gα o −/− hippocampi show no obvious changes in mossy fiber tract anatomy as judged by Synaptoporin stainings. (g–j) Absolute changes are displayed as relative changes from P2 to adulthood. Values for P2 (both for wild‐type and Gα o −/− mice) were set as 1. (g) Fold changes in SLU area. (h) Fold changes in total MFT area. (i) Fold changes in IPB length. (j) Fold changes in total MFT length. Data show means ± SD, N values correspond to the information given in (d, e).

Article Snippet: A polyclonal antibody preferentially recognizing Gα o1 was from Santa Cruz (cat. no. 13532, Santa Cruz, CA, USA).

Techniques: Double Knockout, Knock-Out, Isolation, Amplification, Western Blot, Expressing, Staining, Control, Immunohistochemical staining, Immunostaining

Journal: Journal of Neurochemistry

Article Title: Gα o1 and Gα o1 /Gα o2 deletion differentially affect hippocampal mossy fiber tract anatomy and neuronal morphogenesis

doi: 10.1111/jnc.16248

Figure Lengend Snippet: Knockout of Gα o1 reduces axonal and dendritic length and branching in cultured hippocampal neurons. Morphometrical measurements of cultured hippocampal neurons. (a) Hippocampal neurons were obtained from embryonic wild‐type, Gα o1 +/− , and Gα o1 −/− brains and maintained for 5 days in culture. Following fixation, neurons were stained for Microtubule‐associated protein 2 (Map2) and neurofilament protein (NFP) as dendritic and axonal markers. Morphometrical parameters were analyzed using Neurolucida software. Both heterozygous and homozygous knockout resulted in significant reduction of axonal and dendritic length and branching, more pronounced in the homozygous genotype. In addition, Gα o1 −/− neurons showed a reduction in the number of dendrites developed. *** p ≤ 0.001 (b) Representative images of Map2 and NFP double stainings and alterations of neuronal morphology by the respective knockout types. (c) Sholl analysis of axonal (left) and dendritic (right) growth and complexity. A number of intersections of neuronal branches with circles centered on the soma with 10 μm increment in the distance from the soma were counted. Wild‐type neurons showed a significantly higher number of axon intersections over a broad range of distances from the soma and reached further distances especially than Gα o1 −/− neurons. Heterozygous neurons showed a less pronounced effect. Number of dendritic intersections was also higher in the wild‐type than in knockout neurons, the maximal distance reached was comparable for the three genotypes. Summarized p values indicated by asterisks in the upper rows apply to differences between wild‐type and Gα o1 −/− neurons, asterisks in the lower row indicate differences between wild‐type and Gα o1 +/− neurons. * p ≤ 0.05, ** p ≤ 0.005, *** p ≤ 0.001 (d) Branch order analysis of axonal and dendritic growth. Compared to the wild‐type, both heterozygous and homozygous knockout neurons displayed a reduced development of higher‐order branches (expressed as probability of occurrence in % of neurons). Dendrites at DIV 5 generally show a less complex branching pattern than axons, but, however, also exhibit a reduced expression 2nd‐ to 4th‐order branches. Data in (a), (c) and (d) are given as means ± SEM from three independent experiments, 90 neurons were analyzed per individual condition.

Article Snippet: A polyclonal antibody preferentially recognizing Gα o1 was from Santa Cruz (cat. no. 13532, Santa Cruz, CA, USA).

Techniques: Knock-Out, Cell Culture, Staining, Software, Expressing

Journal: Journal of Neurochemistry

Article Title: Gα o1 and Gα o1 /Gα o2 deletion differentially affect hippocampal mossy fiber tract anatomy and neuronal morphogenesis

doi: 10.1111/jnc.16248

Figure Lengend Snippet: Double knockout of Gα o1 and Gα o1 restores wild‐type axonal and dendritic length and branching in cultured hippocampal neurons. Morphometrical measurements of cultured hippocampal neurons. (a) Hippocampal neurons were obtained from embryonic wild‐type, Gα o +/− and Gα o −/− brains and maintained for 5 days in culture. Following fixation, neurons were stained for Microtubule‐associated protein 2 (Map2) and neurofilament protein (NFP) as dendritic and axonal markers. Morphometrical parameters were analyzed using Neurolucida software. Mean dendritic length was slightly increased in Gα o heterozygous mice, all other dendritic and axonal parameters looked at were unchanged. * p ≤ 0.05 (b) Representative images of Gα o1/2 and Map2 double staining in cultured neurons. Both knockout types result in a loss of immune signal. (c) Representative images of combined Map2 and NFP stainings to demonstrate the similar morphology of all three genotypes. (d) Sholl analysis of axonal (left) and dendritic (right) growth and complexity. A number of intersections of neuronal branches with circles centered on the soma with 10 μm increment in the distance from the soma were counted. No significant changes were observed between wild and Gα o −/− type neurons, and some minor changes were detectable between wild‐type and Gα o +/− neurons. Asterisks apply to differences between wild‐type and Gα o1 +/− neurons. * p ≤ 0.05 (e) Branch order analysis of axonal and dendritic growth. Compared to the wild‐type, both heterozygous and homozygous knockout neurons developed similar axonal and dendritic branch orders. Data in (a), (d) and (e) are given as means ± SEM from three independent experiments, 90 neurons were analyzed per individual condition.

Article Snippet: A polyclonal antibody preferentially recognizing Gα o1 was from Santa Cruz (cat. no. 13532, Santa Cruz, CA, USA).

Techniques: Double Knockout, Cell Culture, Staining, Software, Double Staining, Knock-Out