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Nanjing Jiancheng Bioengineering Research Institute Co Ltd o2 assay kits
O2 Assay Kits, supplied by Nanjing Jiancheng Bioengineering Research Institute Co Ltd, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Colorimetric Assay:

Article Title: PCK2-Mediated PQBP1 Lactylation Promotes Asthmatic Inflammation through PRMT5 Inhibition
Article Snippet: .. The levels of key oxidative stress markers were quantified using commercially available colorimetric assay kits from the Nanjing Jiancheng Bioengineering Institute, including MDA (#A003-1-2), CAT) (#A007-1-1), and SOD (#A001-3-2). .. The levels of key oxidative stress markers were quantified using commercially available colorimetric assay kits from the Nanjing Jiancheng Bioengineering Institute, including MDA (#A003-1-2), CAT) (#A007-1-1), and SOD (#A001-3-2).

Article Title: Dietary thyme essential oil supplementation improves production performance, egg quality, antioxidant status, and intestinal health in late-phase laying hens
Article Snippet: Yolk was separated from albumen using a commercial egg separator (Shanghai Specimen and Model Factory, Shanghai, China), and yolk weight was subsequently recorded. .. Serum antioxidant indices were quantified using commercial colorimetric assay kits (Nanjing Jiancheng Bioengineering Institute, Nanjing, China) according to the manufacturer’s protocols. ..

Multiple Displacement Amplification:

Article Title: PCK2-Mediated PQBP1 Lactylation Promotes Asthmatic Inflammation through PRMT5 Inhibition
Article Snippet: .. The levels of key oxidative stress markers were quantified using commercially available colorimetric assay kits from the Nanjing Jiancheng Bioengineering Institute, including MDA (#A003-1-2), CAT) (#A007-1-1), and SOD (#A001-3-2). .. The levels of key oxidative stress markers were quantified using commercially available colorimetric assay kits from the Nanjing Jiancheng Bioengineering Institute, including MDA (#A003-1-2), CAT) (#A007-1-1), and SOD (#A001-3-2).

Article Title: Semaglutide exerts anti-pulmonary fibrosis effects by inhibiting cellular senescence through activation of the Sirt1/HSF1/HSPs pathway.
Article Snippet: Idiopathic pulmonary fibrosis is an interstitial lung disease of unknown etiology with no effective treatment.. Semaglutide is a glucagon-like peptide-1 receptor agonist that has been widely used in the treatment of diabetes and obesity.. This study investigated the therapeutic effects of semaglutide on pulmonary fibrosis and its molecular mechanisms.

Article Title: Discovery of a bamboo sap-derived dihydroxyacetophenone analog as a potent anti-inflammatory agent against acute liver failure.
Article Snippet: Acute liver failure (ALF) remains a life-threatening condition with few effective therapeutic options.. Bioassayguided fractionation of bamboo sap (Succus Bambusae) enabled identification of 1-(2,3-dihydroxyphenyl) ethan-1-one (ZL-2, IC50 = 17.25 μM) as an anti-inflammatory lead compound.. Synthesis and structure-activity relationship (SAR) evaluation of 27 structural analogs revealed that both the hydroxyl substitution pattern and acyl chain length critically govern anti-inflammatory potency and cytotoxicity.

Article Title: Nao-xin-tong capsule attenuates cerebral ischemia/reperfusion injury by modulating the p53/GDF15/NOX4 regulatory network and alleviating mitochondrial dysfunction and ferroptosis-related injury.
Article Snippet: 21 Ethnopharmacological relevance: Nao-xin-tong (NXT) is a representative traditional Chinese medicine prescription for 22 replenishing qi and activating blood circulation, and has been widely used in the prevention and treatment of 23 cardio-cerebrovascular diseases.. However, the mechanisms underlying its protective effects against cerebral 24 ischemia/reperfusion injury (CIRI) remain incompletely understood.. 25 Aim of the study: This study aimed to investigate whether NXT alleviates CIRI by regulating the p53/GDF15/NOX4 26 regulatory network, thereby improving mitochondrial dysfunction and inhibiting neuronal ferroptosis.

Article Title: Ceritinib induces ferroptosis via TRIM21-mediated GLUT1 ubiquitination and AMPK-driven metabolic reprogramming in breast cancer
Article Snippet: Lipid Peroxidation Assay Kit , Nanjing Jiancheng Bioengineering Institute , Cat# A106-1-1. .. MDA Assay Kit , Nanjing Jiancheng Bioengineering Institute , Cat# A003-4-1. .. Iron Assay Kit , Nanjing Jiancheng Bioengineering Institute , Cat# A039-2-1.

Article Title: Biomimetic macrophage membrane-coated nanoparticles for targeted cardioprotection against myocardial ischaemia-reperfusion injury.
Article Snippet: .. Malondialdehyde (MDA) content and total superoxide dismutase (SOD) activity were then measured using the MDA assay kit (thiobarbituric acid reactive substances (TBARS) method, A003-4-1, Nanjing Jiancheng Bioengineering Institute) and SOD assay kit (A001-1, Nanjing Jiancheng Bioengineering Institute), respectively. .. Absorbance was measured at 450 nm (for all ELISA kits), 532 nm (for MDA), and 550 nm (for SOD) using a microplate reader (Molecular Devices, SpectraMax i3x).

Cholesterol Assay:

Article Title: Targeted activation of PPARG or AKT1 alleviates liver injury in mice with type 2 diabetes and sepsis by modulating inflammatory and metabolic pathways.
Article Snippet: .. The Triglyceride Assay Kit (A110-1-1), the High-Density Lipoprotein Cholesterol Assay Kit (A112-1-1), the Low-Density Lipoprotein Cholesterol Assay Kit (A113-1-1), the Alanine Aminotransferase Assay Kit (C009-11), and the Aspartate Aminotransferase Assay Kit (C010-2-1) (Nanjing Jiancheng, China) were obtained to quantify triglycerides (TG), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), ALT, and AST levels in mouse serum. ..

AST Assay:

Article Title: Targeted activation of PPARG or AKT1 alleviates liver injury in mice with type 2 diabetes and sepsis by modulating inflammatory and metabolic pathways.
Article Snippet: .. The Triglyceride Assay Kit (A110-1-1), the High-Density Lipoprotein Cholesterol Assay Kit (A112-1-1), the Low-Density Lipoprotein Cholesterol Assay Kit (A113-1-1), the Alanine Aminotransferase Assay Kit (C009-11), and the Aspartate Aminotransferase Assay Kit (C010-2-1) (Nanjing Jiancheng, China) were obtained to quantify triglycerides (TG), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), ALT, and AST levels in mouse serum. ..

Activity Assay:

Article Title: Semaglutide exerts anti-pulmonary fibrosis effects by inhibiting cellular senescence through activation of the Sirt1/HSF1/HSPs pathway.
Article Snippet: Idiopathic pulmonary fibrosis is an interstitial lung disease of unknown etiology with no effective treatment.. Semaglutide is a glucagon-like peptide-1 receptor agonist that has been widely used in the treatment of diabetes and obesity.. This study investigated the therapeutic effects of semaglutide on pulmonary fibrosis and its molecular mechanisms.

Article Title: Nao-xin-tong capsule attenuates cerebral ischemia/reperfusion injury by modulating the p53/GDF15/NOX4 regulatory network and alleviating mitochondrial dysfunction and ferroptosis-related injury.
Article Snippet: 21 Ethnopharmacological relevance: Nao-xin-tong (NXT) is a representative traditional Chinese medicine prescription for 22 replenishing qi and activating blood circulation, and has been widely used in the prevention and treatment of 23 cardio-cerebrovascular diseases.. However, the mechanisms underlying its protective effects against cerebral 24 ischemia/reperfusion injury (CIRI) remain incompletely understood.. 25 Aim of the study: This study aimed to investigate whether NXT alleviates CIRI by regulating the p53/GDF15/NOX4 26 regulatory network, thereby improving mitochondrial dysfunction and inhibiting neuronal ferroptosis.

Article Title: Biomimetic macrophage membrane-coated nanoparticles for targeted cardioprotection against myocardial ischaemia-reperfusion injury.
Article Snippet: .. Malondialdehyde (MDA) content and total superoxide dismutase (SOD) activity were then measured using the MDA assay kit (thiobarbituric acid reactive substances (TBARS) method, A003-4-1, Nanjing Jiancheng Bioengineering Institute) and SOD assay kit (A001-1, Nanjing Jiancheng Bioengineering Institute), respectively. .. Absorbance was measured at 450 nm (for all ELISA kits), 532 nm (for MDA), and 550 nm (for SOD) using a microplate reader (Molecular Devices, SpectraMax i3x).

Glutathione Assay:

Article Title: Semaglutide exerts anti-pulmonary fibrosis effects by inhibiting cellular senescence through activation of the Sirt1/HSF1/HSPs pathway.
Article Snippet: Idiopathic pulmonary fibrosis is an interstitial lung disease of unknown etiology with no effective treatment.. Semaglutide is a glucagon-like peptide-1 receptor agonist that has been widely used in the treatment of diabetes and obesity.. This study investigated the therapeutic effects of semaglutide on pulmonary fibrosis and its molecular mechanisms.

Glutathione Peroxidase Assay:

Article Title: Nao-xin-tong capsule attenuates cerebral ischemia/reperfusion injury by modulating the p53/GDF15/NOX4 regulatory network and alleviating mitochondrial dysfunction and ferroptosis-related injury.
Article Snippet: 21 Ethnopharmacological relevance: Nao-xin-tong (NXT) is a representative traditional Chinese medicine prescription for 22 replenishing qi and activating blood circulation, and has been widely used in the prevention and treatment of 23 cardio-cerebrovascular diseases.. However, the mechanisms underlying its protective effects against cerebral 24 ischemia/reperfusion injury (CIRI) remain incompletely understood.. 25 Aim of the study: This study aimed to investigate whether NXT alleviates CIRI by regulating the p53/GDF15/NOX4 26 regulatory network, thereby improving mitochondrial dysfunction and inhibiting neuronal ferroptosis.

ATPase Assay:

Article Title: Nao-xin-tong capsule attenuates cerebral ischemia/reperfusion injury by modulating the p53/GDF15/NOX4 regulatory network and alleviating mitochondrial dysfunction and ferroptosis-related injury.
Article Snippet: 21 Ethnopharmacological relevance: Nao-xin-tong (NXT) is a representative traditional Chinese medicine prescription for 22 replenishing qi and activating blood circulation, and has been widely used in the prevention and treatment of 23 cardio-cerebrovascular diseases.. However, the mechanisms underlying its protective effects against cerebral 24 ischemia/reperfusion injury (CIRI) remain incompletely understood.. 25 Aim of the study: This study aimed to investigate whether NXT alleviates CIRI by regulating the p53/GDF15/NOX4 26 regulatory network, thereby improving mitochondrial dysfunction and inhibiting neuronal ferroptosis.



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<t>Gα</t> o1 and Gα <t>o2</t> differentially affect the number of VGLUT2 contacts within the cerebellar cortex. (A) Immunohistochemical analysis of the vesicular glutamate transporter 2 (VGLUT2) expression in the adult wild type cerebellum as shown in a midsagittal section. A strong immune signal was detected in the cerebellar glomeruli of the granule cell layer (gcl), together with a distinct punctuate pattern in the molecular layer (ml) and the deep cerebellar nuclei (DCN, see inset for Dapi staining). (B) Double staining for VGLUT2 and Calbindin to visualize Purkinje cell dendrite morphology. In the molecular layer, the VGLUT2 transporter signal characteristically marks synaptic climbing fiber contacts to the proximal to mid‐segmental Purkinje cell dendrites. (C) Regions of the molecular layer depicting VGLUT2 signals in the culmen of wild type and Gα o1 −/− mice used to quantify the VGLUT2 puncta. (D) Exemplary thresholded images of the regions shown in (C) used for analysis in the respective regions of interest (counting area, shown in transparent blue, see insets for magnification). (E) Quantification of the absolute number of VGLUT2 puncta in the counting area (left panel), the density (middle panel), and size (right panel). See higher magnification confocal imaging for details of VGLUT2 signals. (F) Analysis of the effects of Gα o2 knockout on the VGLUT2 staining pattern in the molecular layer of the cerebellum. Exemplary immunohistochemical visualization of VGLUT2 contacts in the central lobule II of wild type and Gα o2 −/− knockout mice. (G) Exemplary thresholded images of the regions used for quantification within the counting area (transparent blue). (H) Quantification of the absolute number of VGLUT2 puncta in the counting area (left panel), the density (middle panel), and size (right panel). See higher magnification confocal imaging for details of VGLUT2 signals. (I) Analysis of the effects of Gα o1/2 (Gα o ) double knockout on the VGLUT2 staining pattern in the molecular layer of the cerebellum. Exemplary immunohistochemical visualization of VGLUT2 contacts in the declive of wild type and Gα o −/− knockout mice. (J) Exemplary thresholded images of the above depicted regions used for quantification within the counting area (transparent blue). (K) Quantification of the absolute number of VGLUT2 puncta in the counting area (left panel), the density (middle panel), and size (right panel). See higher magnification confocal imaging for details of VGLUT2 signals. Bars show means ± SD (individual animal means are depicted), N = 4 animals each WT and knockout, 6–8 sections per animal and genotype. *** p ≤ 0.001.
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<t>Gα</t> o1 and Gα <t>o2</t> differentially affect the number of VGLUT2 contacts within the cerebellar cortex. (A) Immunohistochemical analysis of the vesicular glutamate transporter 2 (VGLUT2) expression in the adult wild type cerebellum as shown in a midsagittal section. A strong immune signal was detected in the cerebellar glomeruli of the granule cell layer (gcl), together with a distinct punctuate pattern in the molecular layer (ml) and the deep cerebellar nuclei (DCN, see inset for Dapi staining). (B) Double staining for VGLUT2 and Calbindin to visualize Purkinje cell dendrite morphology. In the molecular layer, the VGLUT2 transporter signal characteristically marks synaptic climbing fiber contacts to the proximal to mid‐segmental Purkinje cell dendrites. (C) Regions of the molecular layer depicting VGLUT2 signals in the culmen of wild type and Gα o1 −/− mice used to quantify the VGLUT2 puncta. (D) Exemplary thresholded images of the regions shown in (C) used for analysis in the respective regions of interest (counting area, shown in transparent blue, see insets for magnification). (E) Quantification of the absolute number of VGLUT2 puncta in the counting area (left panel), the density (middle panel), and size (right panel). See higher magnification confocal imaging for details of VGLUT2 signals. (F) Analysis of the effects of Gα o2 knockout on the VGLUT2 staining pattern in the molecular layer of the cerebellum. Exemplary immunohistochemical visualization of VGLUT2 contacts in the central lobule II of wild type and Gα o2 −/− knockout mice. (G) Exemplary thresholded images of the regions used for quantification within the counting area (transparent blue). (H) Quantification of the absolute number of VGLUT2 puncta in the counting area (left panel), the density (middle panel), and size (right panel). See higher magnification confocal imaging for details of VGLUT2 signals. (I) Analysis of the effects of Gα o1/2 (Gα o ) double knockout on the VGLUT2 staining pattern in the molecular layer of the cerebellum. Exemplary immunohistochemical visualization of VGLUT2 contacts in the declive of wild type and Gα o −/− knockout mice. (J) Exemplary thresholded images of the above depicted regions used for quantification within the counting area (transparent blue). (K) Quantification of the absolute number of VGLUT2 puncta in the counting area (left panel), the density (middle panel), and size (right panel). See higher magnification confocal imaging for details of VGLUT2 signals. Bars show means ± SD (individual animal means are depicted), N = 4 animals each WT and knockout, 6–8 sections per animal and genotype. *** p ≤ 0.001.
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Image Search Results


Gα o1 and Gα o2 differentially affect the number of VGLUT2 contacts within the cerebellar cortex. (A) Immunohistochemical analysis of the vesicular glutamate transporter 2 (VGLUT2) expression in the adult wild type cerebellum as shown in a midsagittal section. A strong immune signal was detected in the cerebellar glomeruli of the granule cell layer (gcl), together with a distinct punctuate pattern in the molecular layer (ml) and the deep cerebellar nuclei (DCN, see inset for Dapi staining). (B) Double staining for VGLUT2 and Calbindin to visualize Purkinje cell dendrite morphology. In the molecular layer, the VGLUT2 transporter signal characteristically marks synaptic climbing fiber contacts to the proximal to mid‐segmental Purkinje cell dendrites. (C) Regions of the molecular layer depicting VGLUT2 signals in the culmen of wild type and Gα o1 −/− mice used to quantify the VGLUT2 puncta. (D) Exemplary thresholded images of the regions shown in (C) used for analysis in the respective regions of interest (counting area, shown in transparent blue, see insets for magnification). (E) Quantification of the absolute number of VGLUT2 puncta in the counting area (left panel), the density (middle panel), and size (right panel). See higher magnification confocal imaging for details of VGLUT2 signals. (F) Analysis of the effects of Gα o2 knockout on the VGLUT2 staining pattern in the molecular layer of the cerebellum. Exemplary immunohistochemical visualization of VGLUT2 contacts in the central lobule II of wild type and Gα o2 −/− knockout mice. (G) Exemplary thresholded images of the regions used for quantification within the counting area (transparent blue). (H) Quantification of the absolute number of VGLUT2 puncta in the counting area (left panel), the density (middle panel), and size (right panel). See higher magnification confocal imaging for details of VGLUT2 signals. (I) Analysis of the effects of Gα o1/2 (Gα o ) double knockout on the VGLUT2 staining pattern in the molecular layer of the cerebellum. Exemplary immunohistochemical visualization of VGLUT2 contacts in the declive of wild type and Gα o −/− knockout mice. (J) Exemplary thresholded images of the above depicted regions used for quantification within the counting area (transparent blue). (K) Quantification of the absolute number of VGLUT2 puncta in the counting area (left panel), the density (middle panel), and size (right panel). See higher magnification confocal imaging for details of VGLUT2 signals. Bars show means ± SD (individual animal means are depicted), N = 4 animals each WT and knockout, 6–8 sections per animal and genotype. *** p ≤ 0.001.

Journal: Journal of Neurochemistry

Article Title: Splice Type‐Specific Effects of Gαo Subunits on Cerebellar Anatomy and Synapse Formation

doi: 10.1111/jnc.70512

Figure Lengend Snippet: Gα o1 and Gα o2 differentially affect the number of VGLUT2 contacts within the cerebellar cortex. (A) Immunohistochemical analysis of the vesicular glutamate transporter 2 (VGLUT2) expression in the adult wild type cerebellum as shown in a midsagittal section. A strong immune signal was detected in the cerebellar glomeruli of the granule cell layer (gcl), together with a distinct punctuate pattern in the molecular layer (ml) and the deep cerebellar nuclei (DCN, see inset for Dapi staining). (B) Double staining for VGLUT2 and Calbindin to visualize Purkinje cell dendrite morphology. In the molecular layer, the VGLUT2 transporter signal characteristically marks synaptic climbing fiber contacts to the proximal to mid‐segmental Purkinje cell dendrites. (C) Regions of the molecular layer depicting VGLUT2 signals in the culmen of wild type and Gα o1 −/− mice used to quantify the VGLUT2 puncta. (D) Exemplary thresholded images of the regions shown in (C) used for analysis in the respective regions of interest (counting area, shown in transparent blue, see insets for magnification). (E) Quantification of the absolute number of VGLUT2 puncta in the counting area (left panel), the density (middle panel), and size (right panel). See higher magnification confocal imaging for details of VGLUT2 signals. (F) Analysis of the effects of Gα o2 knockout on the VGLUT2 staining pattern in the molecular layer of the cerebellum. Exemplary immunohistochemical visualization of VGLUT2 contacts in the central lobule II of wild type and Gα o2 −/− knockout mice. (G) Exemplary thresholded images of the regions used for quantification within the counting area (transparent blue). (H) Quantification of the absolute number of VGLUT2 puncta in the counting area (left panel), the density (middle panel), and size (right panel). See higher magnification confocal imaging for details of VGLUT2 signals. (I) Analysis of the effects of Gα o1/2 (Gα o ) double knockout on the VGLUT2 staining pattern in the molecular layer of the cerebellum. Exemplary immunohistochemical visualization of VGLUT2 contacts in the declive of wild type and Gα o −/− knockout mice. (J) Exemplary thresholded images of the above depicted regions used for quantification within the counting area (transparent blue). (K) Quantification of the absolute number of VGLUT2 puncta in the counting area (left panel), the density (middle panel), and size (right panel). See higher magnification confocal imaging for details of VGLUT2 signals. Bars show means ± SD (individual animal means are depicted), N = 4 animals each WT and knockout, 6–8 sections per animal and genotype. *** p ≤ 0.001.

Article Snippet: Monoclonal antibodies specifically recognizing Gα o2 were from Synaptic Systems (cat. no. 271011, clone 101.4, Western blot) and 201.2 (kind gift from Karsten Spicher, see Spicher et al. , immunofluorescence).

Techniques: Immunohistochemical staining, Expressing, Staining, Double Staining, Imaging, Knock-Out, Double Knockout

Gα o1 and Gα o2 differentially affect the number of VGLUT1 contacts within the cerebellar cortex. Immunohistochemical analysis of the vesicular glutamate transporter 1 (VGLUT1) expression in the adult wild type cerebellum as shown in a midsagittal section. A very dense immune signal was detected in the molecular layer. In addition, the cerebellar glomeruli of the granule cell layer (gcl), were marked (see inset for Dapi staining). (B) Double staining for VGLUT1 and Calbindin. In the molecular layer, the VGLUT1 transporter signal forms a dense synaptic carpet sparing the main Purkinje cell dendrite branches. (C) Expression of VGLUT1 in the individual folia. Cerebellar wild type and Gα o1 −/− sections were incubated with a VGLUT1 antibody and protein expression was quantified for the molecular layer of the individual lobules and expressed as brightness values. See representative images. Scale bar 50 μm. Expression levels of VGLUT1 averaged over all folia are given in the bar chart. (D) The same analysis was performed for wild type and Gα o2 −/− cerebella. See representative images. Scale bar 50 μm. Expression levels of VGLUT1 averaged over all folia are given in the bar chart. (E) Expression of VGLUT1 in the individual folia of wild type and Gα o −/− cerebella. See representative images. Scale bar 50 μm. Expression levels of VGLUT1 averaged over all folia are given in the bar chart. (F) Analysis of the effects of Gα o1 knockout on the VGLUT1 staining pattern in the central lobule II of wild type and Gα o1 −/− mice used to quantify individual VGLUT1 puncta. Exemplary thresholded images are shown used for analysis in the respective regions of interest (counting area, shown in transparent blue). (G) Analysis of the effects of Gα o2 knockout on the VGLUT1 staining pattern in the uvula of wild type and Gα o2 −/− mice as shown in (F). (H) Analysis of the effects of Gα o knockout on the VGLUT1 staining pattern in the declive of wild type and Gα o −/− mice as shown in (F) and (G). (B, F–H) Confocal imaging. Data show means ± SD with individual animal means or individual sections depicted in bar charts, N = 3 animals each WT and knockout. 5–6 sections per animal and genotype. * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001.

Journal: Journal of Neurochemistry

Article Title: Splice Type‐Specific Effects of Gαo Subunits on Cerebellar Anatomy and Synapse Formation

doi: 10.1111/jnc.70512

Figure Lengend Snippet: Gα o1 and Gα o2 differentially affect the number of VGLUT1 contacts within the cerebellar cortex. Immunohistochemical analysis of the vesicular glutamate transporter 1 (VGLUT1) expression in the adult wild type cerebellum as shown in a midsagittal section. A very dense immune signal was detected in the molecular layer. In addition, the cerebellar glomeruli of the granule cell layer (gcl), were marked (see inset for Dapi staining). (B) Double staining for VGLUT1 and Calbindin. In the molecular layer, the VGLUT1 transporter signal forms a dense synaptic carpet sparing the main Purkinje cell dendrite branches. (C) Expression of VGLUT1 in the individual folia. Cerebellar wild type and Gα o1 −/− sections were incubated with a VGLUT1 antibody and protein expression was quantified for the molecular layer of the individual lobules and expressed as brightness values. See representative images. Scale bar 50 μm. Expression levels of VGLUT1 averaged over all folia are given in the bar chart. (D) The same analysis was performed for wild type and Gα o2 −/− cerebella. See representative images. Scale bar 50 μm. Expression levels of VGLUT1 averaged over all folia are given in the bar chart. (E) Expression of VGLUT1 in the individual folia of wild type and Gα o −/− cerebella. See representative images. Scale bar 50 μm. Expression levels of VGLUT1 averaged over all folia are given in the bar chart. (F) Analysis of the effects of Gα o1 knockout on the VGLUT1 staining pattern in the central lobule II of wild type and Gα o1 −/− mice used to quantify individual VGLUT1 puncta. Exemplary thresholded images are shown used for analysis in the respective regions of interest (counting area, shown in transparent blue). (G) Analysis of the effects of Gα o2 knockout on the VGLUT1 staining pattern in the uvula of wild type and Gα o2 −/− mice as shown in (F). (H) Analysis of the effects of Gα o knockout on the VGLUT1 staining pattern in the declive of wild type and Gα o −/− mice as shown in (F) and (G). (B, F–H) Confocal imaging. Data show means ± SD with individual animal means or individual sections depicted in bar charts, N = 3 animals each WT and knockout. 5–6 sections per animal and genotype. * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001.

Article Snippet: Monoclonal antibodies specifically recognizing Gα o2 were from Synaptic Systems (cat. no. 271011, clone 101.4, Western blot) and 201.2 (kind gift from Karsten Spicher, see Spicher et al. , immunofluorescence).

Techniques: Immunohistochemical staining, Expressing, Staining, Double Staining, Incubation, Knock-Out, Imaging

Gα o1 and Gα o2 differentially affect the cerebellar size. (A) Immunohistochemical analysis of Gα o1 expression in the cerebellum of wild type and Gα o1 −/− mice. Midsagittal cerebellum sections of adult mice of either strain were stained by an antibody preferentially recognizing Gα o1 . In the wild type, Gα o1 showed a very strong reaction predominantly in the molecular layer. Immunostaining against Gα o1 was absent in the knockout. gcl, granule cell layer; ml, molecular layer; pcl, Purkinje cell layer. (B) Analysis of Gα o2 expression. Midsagittal cerebellum sections of adult mice were stained by an antibody recognizing Gα o2 . In the wild type, Gα o2 showed a strong reaction predominantly in the molecular layer. Immunostaining against Gα o2 was absent in the knockout. (C) Analysis of Gα o1/2 (Gα o ) expression. Midsagittal cerebellum sections of adult mice of either strain were stained by an antibody recognizing both Gα o1 and Gα o2 . In the wild type, Gα o showed a very strong reaction predominantly in the molecular layer. Immunostaining against Gα o was absent in the knockout. (D) An exemplary midsagittal DAPI‐stained section of wild type cerebellum with the individual folia labeled. Cranial orientation is to the left. CENT, central lobule; CUL, culmen; DEC, declive; FOTU, fotulum‐tuber‐vermis; NOD, nodulus; PYR, pyramis; UVU, uvula (lobule numbers are given in latin). (E) Comparison of wild type (as shown in D) and Gα o1 −/− cerebellum size. (F) Comparison of wild type and Gα o2 −/− cerebellum size. (G) Comparison of wild type and Gα o −/− cerebellum size. (H) Quantification of overall cerebellar size (given in mm 2 , upper left panel) along with the cranio‐caudal (length) and ventro‐dorsal (height) extension (lower left panel). In addition, the cortical area was calculated for the molecular and granule cell layer (lower right panel). See effects exemplified for CENTIII by Calbindin staining. (I) The same analysis was performed for Gα o2 . See effects exemplified for CENTII by Calbindin staining, (J) Analysis for Gα o . See effects exemplified for CENTII by Calbindin staining. Bars show means ± SD and individual values, N = 4 animals each WT and knockout (except for 3 animals in Gα o2 WT total ML and GCL area analysis), 6–8 sections per animal and genotype. * p ≤ 0.05; ** p ≤ 0.01; *** p ≤ 0.001.

Journal: Journal of Neurochemistry

Article Title: Splice Type‐Specific Effects of Gαo Subunits on Cerebellar Anatomy and Synapse Formation

doi: 10.1111/jnc.70512

Figure Lengend Snippet: Gα o1 and Gα o2 differentially affect the cerebellar size. (A) Immunohistochemical analysis of Gα o1 expression in the cerebellum of wild type and Gα o1 −/− mice. Midsagittal cerebellum sections of adult mice of either strain were stained by an antibody preferentially recognizing Gα o1 . In the wild type, Gα o1 showed a very strong reaction predominantly in the molecular layer. Immunostaining against Gα o1 was absent in the knockout. gcl, granule cell layer; ml, molecular layer; pcl, Purkinje cell layer. (B) Analysis of Gα o2 expression. Midsagittal cerebellum sections of adult mice were stained by an antibody recognizing Gα o2 . In the wild type, Gα o2 showed a strong reaction predominantly in the molecular layer. Immunostaining against Gα o2 was absent in the knockout. (C) Analysis of Gα o1/2 (Gα o ) expression. Midsagittal cerebellum sections of adult mice of either strain were stained by an antibody recognizing both Gα o1 and Gα o2 . In the wild type, Gα o showed a very strong reaction predominantly in the molecular layer. Immunostaining against Gα o was absent in the knockout. (D) An exemplary midsagittal DAPI‐stained section of wild type cerebellum with the individual folia labeled. Cranial orientation is to the left. CENT, central lobule; CUL, culmen; DEC, declive; FOTU, fotulum‐tuber‐vermis; NOD, nodulus; PYR, pyramis; UVU, uvula (lobule numbers are given in latin). (E) Comparison of wild type (as shown in D) and Gα o1 −/− cerebellum size. (F) Comparison of wild type and Gα o2 −/− cerebellum size. (G) Comparison of wild type and Gα o −/− cerebellum size. (H) Quantification of overall cerebellar size (given in mm 2 , upper left panel) along with the cranio‐caudal (length) and ventro‐dorsal (height) extension (lower left panel). In addition, the cortical area was calculated for the molecular and granule cell layer (lower right panel). See effects exemplified for CENTIII by Calbindin staining. (I) The same analysis was performed for Gα o2 . See effects exemplified for CENTII by Calbindin staining, (J) Analysis for Gα o . See effects exemplified for CENTII by Calbindin staining. Bars show means ± SD and individual values, N = 4 animals each WT and knockout (except for 3 animals in Gα o2 WT total ML and GCL area analysis), 6–8 sections per animal and genotype. * p ≤ 0.05; ** p ≤ 0.01; *** p ≤ 0.001.

Article Snippet: Monoclonal antibodies specifically recognizing Gα o2 were from Synaptic Systems (cat. no. 271011, clone 101.4, Western blot) and 201.2 (kind gift from Karsten Spicher, see Spicher et al. , immunofluorescence).

Techniques: Immunohistochemical staining, Expressing, Staining, Immunostaining, Knock-Out, Labeling, Comparison

Folia‐specific Gαo subunit expression and determination of Gα o1 and Gα o2 knockout effects on VGLUT2‐positive synaptic contacts within the cerebellar cortex. Displayed are the effects of Gα o1 , Gα o2 , and Gα o knockout on the VGLUT2‐positive synaptic contacts in the molecular layer of the individual folia of the vermis. The same analysis as summarized in Figure for the total molecular layer was conducted for the molecular layer at the level of individual folia. (A) Folia‐specific Gα o1 knockout effects were observed for the absolute number (left panel), density (middle panel) and size of VGLUT2 puncta (right panel). (B) Folia‐specific Gα o2 knockout effects on the same parameters of VGLUT2 contacts as shown in (A). (C) Folia‐specific Gα o2 knockout effects on the same parameters of VGLUT2 contacts as shown in (A). (D) Expression of Gα o1 in the individual folia. Cerebellar wild type sections were incubated with the Gα o1 antibody and protein expression was quantified for the molecular layer of the individual lobules. The knockout is shown for control. See representative images. (E) The same analysis was performed for the expression of Gα o2 . See representative images. (F) Analysis of the combined expression of both isoforms by the Gα o antibody confirmed the results observed with the subtype‐specific antibodies. See representative images. Data show means ± SD, N = 3–4 animals each WT and knockout, 6–8 sections per animal and genotype. * p ≤ 0.05; ** p ≤ 0.01; *** p ≤ 0.001.

Journal: Journal of Neurochemistry

Article Title: Splice Type‐Specific Effects of Gαo Subunits on Cerebellar Anatomy and Synapse Formation

doi: 10.1111/jnc.70512

Figure Lengend Snippet: Folia‐specific Gαo subunit expression and determination of Gα o1 and Gα o2 knockout effects on VGLUT2‐positive synaptic contacts within the cerebellar cortex. Displayed are the effects of Gα o1 , Gα o2 , and Gα o knockout on the VGLUT2‐positive synaptic contacts in the molecular layer of the individual folia of the vermis. The same analysis as summarized in Figure for the total molecular layer was conducted for the molecular layer at the level of individual folia. (A) Folia‐specific Gα o1 knockout effects were observed for the absolute number (left panel), density (middle panel) and size of VGLUT2 puncta (right panel). (B) Folia‐specific Gα o2 knockout effects on the same parameters of VGLUT2 contacts as shown in (A). (C) Folia‐specific Gα o2 knockout effects on the same parameters of VGLUT2 contacts as shown in (A). (D) Expression of Gα o1 in the individual folia. Cerebellar wild type sections were incubated with the Gα o1 antibody and protein expression was quantified for the molecular layer of the individual lobules. The knockout is shown for control. See representative images. (E) The same analysis was performed for the expression of Gα o2 . See representative images. (F) Analysis of the combined expression of both isoforms by the Gα o antibody confirmed the results observed with the subtype‐specific antibodies. See representative images. Data show means ± SD, N = 3–4 animals each WT and knockout, 6–8 sections per animal and genotype. * p ≤ 0.05; ** p ≤ 0.01; *** p ≤ 0.001.

Article Snippet: Monoclonal antibodies specifically recognizing Gα o2 were from Synaptic Systems (cat. no. 271011, clone 101.4, Western blot) and 201.2 (kind gift from Karsten Spicher, see Spicher et al. , immunofluorescence).

Techniques: Expressing, Knock-Out, Incubation, Control

Differential effects of single Gα o1 , Gα o2 , and double Gα o1 /Gα o2 knockout on the number and size of VGAT‐positive synaptic contacts in the molecular layer. (A) Immunohistochemical analysis of the vesicular GABA transporter (VGAT) expression in the adult wild type cerebellum as shown in a midsagittal section. A strong immune signal was detected at the Purkinje cell somata, in the cerebellar glomeruli of the granule cell layer (gcl), together with a punctuate pattern in the molecular layer (ml) and the deep cerebellar nuclei (DCN, see inset for Dapi staining). (B) Double staining for Calbindin to visualize Purkinje cell dendrite morphology and VGAT. In the molecular layer, the VGAT signal is detected on all segments of the Purkinje cell dendrites. (C) Analysis of the effects of Gα o1 knockout on the VGAT staining pattern in the molecular layer of the cerebellum. Regions of the molecular layer depicting VGAT signals in the central lobule II of wild type and Gα o1 −/− mice were used to quantify the VGAT puncta. (D) Exemplary thresholded images of the regions are shown used for analysis in the respective regions of interest (counting area, shown in transparent blue, see insets for magnification). (E) Quantification of VGAT puncta in the counting area (left panel), their density (middle panel), and size (right panel). See higher magnification confocal imaging for details of VGAT signals. (F) Analysis of the effects of Gα o2 knockout on the VGAT staining pattern in the molecular layer of the cerebellum. Regions of the molecular layer depicting VGAT signals in the central lobule III of wild type and Gα o2 −/− mice were used to quantify the VGAT puncta. (G) Exemplary thresholded images of the regions are shown used for analysis in the respective regions of interest (counting area, shown in transparent blue, see insets for magnification). (H) Quantification of VGAT puncta in the counting area (left panel), density (middle panel), and size (right panel). See higher magnification confocal imaging for details of VGAT signals. (I) Analysis of the effects of Gα o1 and Gα o2 double knockout on the VGAT staining pattern in the molecular layer of the cerebellum. Regions of the molecular layer depicting VGAT signals in the culmen of wild type and Gα o −/− mice were used to quantify the VGAT puncta. (J) Exemplary thresholded images of the regions are shown used for analysis in the respective regions of interest (counting area, shown in transparent blue, see insets for magnification). (K) Quantification of VGAT puncta in the counting area (left panel), their density (middle panel), and size (right panel). See higher magnification confocal imaging for details of VGAT signals. Bars show means ± SD with individual animal means depicted, N = 3 animals each WT/Gα o1 KO as well as WT/Gα o2 KO, N = 4 animals each WT/Gα o2 KO; 6–8 sections per animal and genotype. *** p ≤ 0.001.

Journal: Journal of Neurochemistry

Article Title: Splice Type‐Specific Effects of Gαo Subunits on Cerebellar Anatomy and Synapse Formation

doi: 10.1111/jnc.70512

Figure Lengend Snippet: Differential effects of single Gα o1 , Gα o2 , and double Gα o1 /Gα o2 knockout on the number and size of VGAT‐positive synaptic contacts in the molecular layer. (A) Immunohistochemical analysis of the vesicular GABA transporter (VGAT) expression in the adult wild type cerebellum as shown in a midsagittal section. A strong immune signal was detected at the Purkinje cell somata, in the cerebellar glomeruli of the granule cell layer (gcl), together with a punctuate pattern in the molecular layer (ml) and the deep cerebellar nuclei (DCN, see inset for Dapi staining). (B) Double staining for Calbindin to visualize Purkinje cell dendrite morphology and VGAT. In the molecular layer, the VGAT signal is detected on all segments of the Purkinje cell dendrites. (C) Analysis of the effects of Gα o1 knockout on the VGAT staining pattern in the molecular layer of the cerebellum. Regions of the molecular layer depicting VGAT signals in the central lobule II of wild type and Gα o1 −/− mice were used to quantify the VGAT puncta. (D) Exemplary thresholded images of the regions are shown used for analysis in the respective regions of interest (counting area, shown in transparent blue, see insets for magnification). (E) Quantification of VGAT puncta in the counting area (left panel), their density (middle panel), and size (right panel). See higher magnification confocal imaging for details of VGAT signals. (F) Analysis of the effects of Gα o2 knockout on the VGAT staining pattern in the molecular layer of the cerebellum. Regions of the molecular layer depicting VGAT signals in the central lobule III of wild type and Gα o2 −/− mice were used to quantify the VGAT puncta. (G) Exemplary thresholded images of the regions are shown used for analysis in the respective regions of interest (counting area, shown in transparent blue, see insets for magnification). (H) Quantification of VGAT puncta in the counting area (left panel), density (middle panel), and size (right panel). See higher magnification confocal imaging for details of VGAT signals. (I) Analysis of the effects of Gα o1 and Gα o2 double knockout on the VGAT staining pattern in the molecular layer of the cerebellum. Regions of the molecular layer depicting VGAT signals in the culmen of wild type and Gα o −/− mice were used to quantify the VGAT puncta. (J) Exemplary thresholded images of the regions are shown used for analysis in the respective regions of interest (counting area, shown in transparent blue, see insets for magnification). (K) Quantification of VGAT puncta in the counting area (left panel), their density (middle panel), and size (right panel). See higher magnification confocal imaging for details of VGAT signals. Bars show means ± SD with individual animal means depicted, N = 3 animals each WT/Gα o1 KO as well as WT/Gα o2 KO, N = 4 animals each WT/Gα o2 KO; 6–8 sections per animal and genotype. *** p ≤ 0.001.

Article Snippet: Monoclonal antibodies specifically recognizing Gα o2 were from Synaptic Systems (cat. no. 271011, clone 101.4, Western blot) and 201.2 (kind gift from Karsten Spicher, see Spicher et al. , immunofluorescence).

Techniques: Knock-Out, Immunohistochemical staining, Expressing, Staining, Double Staining, Imaging, Double Knockout

Folia‐specific determination of Gα o1 and Gα o2 knockout effects on VGAT‐positive synaptic contacts within the cerebellar cortex. Displayed are the effects of Gα o1 , Gα o2 , and Gα o knockout on the VGAT‐positive synaptic contacts in the molecular layer of the individual folia of the vermis. (A) Significant reductions appearing in the Gα o1 knockout regarding the absolute number (left panel), density (middle panel), and the size of VGAT puncta (right panel) are indicated. (B) The same analysis was performed for the Gα o2 knockout. Mainly positive significant effects are indicated. (C) Folia‐specific analysis as in (A) and (B) for the Gα o knockout. Significant reductions are indicated. (D) Number of Parvalbumin‐positive cells within the molecular layer of the lobules CENTIII and uvula in wild type and Gα o1 knockout animals. (E) Number of Parvalbumin‐positive cells within the molecular layer of the lobules CENTIII and pyramis in wild type and Gα o2 knockout animals. (F) Number of Parvalbumin‐positive cells within the molecular layer of the lobules CENTIII and culmen in wild type and Gα o knockout animals. See representative images for wild type and Gα o2 −/− cerebella stained with an antibody recognizing Parvalbumin. Asterisks depict cell somata. Data show means ± SD, N = 3 animals each WT/Gα o1 KO as well as WT/Gα o2 KO, N = 4 animals each WT/Gα o2 KO (VGAT analysis). Parvalbumin counting N = 3 animals each condition. * p ≤ 0.05; ** p ≤ 0.01; *** p ≤ 0.001.

Journal: Journal of Neurochemistry

Article Title: Splice Type‐Specific Effects of Gαo Subunits on Cerebellar Anatomy and Synapse Formation

doi: 10.1111/jnc.70512

Figure Lengend Snippet: Folia‐specific determination of Gα o1 and Gα o2 knockout effects on VGAT‐positive synaptic contacts within the cerebellar cortex. Displayed are the effects of Gα o1 , Gα o2 , and Gα o knockout on the VGAT‐positive synaptic contacts in the molecular layer of the individual folia of the vermis. (A) Significant reductions appearing in the Gα o1 knockout regarding the absolute number (left panel), density (middle panel), and the size of VGAT puncta (right panel) are indicated. (B) The same analysis was performed for the Gα o2 knockout. Mainly positive significant effects are indicated. (C) Folia‐specific analysis as in (A) and (B) for the Gα o knockout. Significant reductions are indicated. (D) Number of Parvalbumin‐positive cells within the molecular layer of the lobules CENTIII and uvula in wild type and Gα o1 knockout animals. (E) Number of Parvalbumin‐positive cells within the molecular layer of the lobules CENTIII and pyramis in wild type and Gα o2 knockout animals. (F) Number of Parvalbumin‐positive cells within the molecular layer of the lobules CENTIII and culmen in wild type and Gα o knockout animals. See representative images for wild type and Gα o2 −/− cerebella stained with an antibody recognizing Parvalbumin. Asterisks depict cell somata. Data show means ± SD, N = 3 animals each WT/Gα o1 KO as well as WT/Gα o2 KO, N = 4 animals each WT/Gα o2 KO (VGAT analysis). Parvalbumin counting N = 3 animals each condition. * p ≤ 0.05; ** p ≤ 0.01; *** p ≤ 0.001.

Article Snippet: Monoclonal antibodies specifically recognizing Gα o2 were from Synaptic Systems (cat. no. 271011, clone 101.4, Western blot) and 201.2 (kind gift from Karsten Spicher, see Spicher et al. , immunofluorescence).

Techniques: Knock-Out, Staining

Alterations of Purkinje cell morphology by the knockout of Gα o1 , Gα o2 , and the double knockout. (A) Morphometrical analysis of Purkinje cell primary dendrite width in the CENTIII lobule of wild type and Gα o1 −/− mice. Gα o1 knockout effects are displayed as relative cumulative frequency and overall means. White bars in images represent 3 μm. (B) Analysis as shown in (A) performed for primary dendrites in the culmen. Knockout effects are displayed as above. White bars in images represent 3 μm. (C) Analysis of dendrite outgrowth. Regions of interest (ROI) of 180 × 100 μm within the molecular layer of wild type and Gα o1 −/− cerebella were analyzed and the total Calbindin stained area as well as the proportion of the ROI occupied by the staining were quantified. Thresholded images. (D) Analysis as shown above for the knockout of Gα o2 in the CENTIII lobule. White bars in images represent 3 μm. (E) Analysis of effects in the uvula. White bars in images represent 3 μm. (F) Analysis of dendrite outgrowth as performed in (C). Thresholded images. (G) Analysis performed for primary dendrites in the CENTIII lobule of wild type and Gα o knockout mice. (H) Analysis of knockout effects in the declive. (I) Analysis of dendrite outgrowth as performed in (C) and (F). Thresholded images. Data show means ± SD with individual sections depicted in bar charts, N = 3 animals each WT and knockout. 5–6 sections per animal and genotype. * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001.

Journal: Journal of Neurochemistry

Article Title: Splice Type‐Specific Effects of Gαo Subunits on Cerebellar Anatomy and Synapse Formation

doi: 10.1111/jnc.70512

Figure Lengend Snippet: Alterations of Purkinje cell morphology by the knockout of Gα o1 , Gα o2 , and the double knockout. (A) Morphometrical analysis of Purkinje cell primary dendrite width in the CENTIII lobule of wild type and Gα o1 −/− mice. Gα o1 knockout effects are displayed as relative cumulative frequency and overall means. White bars in images represent 3 μm. (B) Analysis as shown in (A) performed for primary dendrites in the culmen. Knockout effects are displayed as above. White bars in images represent 3 μm. (C) Analysis of dendrite outgrowth. Regions of interest (ROI) of 180 × 100 μm within the molecular layer of wild type and Gα o1 −/− cerebella were analyzed and the total Calbindin stained area as well as the proportion of the ROI occupied by the staining were quantified. Thresholded images. (D) Analysis as shown above for the knockout of Gα o2 in the CENTIII lobule. White bars in images represent 3 μm. (E) Analysis of effects in the uvula. White bars in images represent 3 μm. (F) Analysis of dendrite outgrowth as performed in (C). Thresholded images. (G) Analysis performed for primary dendrites in the CENTIII lobule of wild type and Gα o knockout mice. (H) Analysis of knockout effects in the declive. (I) Analysis of dendrite outgrowth as performed in (C) and (F). Thresholded images. Data show means ± SD with individual sections depicted in bar charts, N = 3 animals each WT and knockout. 5–6 sections per animal and genotype. * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001.

Article Snippet: Monoclonal antibodies specifically recognizing Gα o2 were from Synaptic Systems (cat. no. 271011, clone 101.4, Western blot) and 201.2 (kind gift from Karsten Spicher, see Spicher et al. , immunofluorescence).

Techniques: Knock-Out, Double Knockout, Staining