Journal: Experimental Biology and Medicine

Article Title: Feature article: Splenectomy fails to attenuate immuno-hematologic changes after rodent vertical sleeve gastrectomy

doi: 10.1177/1535370219857991

Figure Lengend Snippet: Spleen gene qPCR.

Article Snippet: * P < 0.05, ** P < 0.01. table ft1 table-wrap mode="anchored" t5 Table 1. caption a7 Sham VSG Gene Gene symbol Probe # Mean±SEM Mean±SEM P value Il6 Interleukin 6 Rn01410330_m1 100±31.8 124.50±24.6 0.493 Il1β Interleukin 1β Rn00580432_m1 100±11.1 83.18±21.9 0.663 TNFα Tumor necrosis factor α Rn01525859_g1 100±20.5 141.60±20.9 0.225 CD68 Cluster of differentiation 68 Rn01495634_g1 100±28.0 73.40±14.5 0.095 HIF1α Hypoxia inducible factor 1α Rn01472831_m1 100±17.6 89.15±16.8 0.796 CASP3 Caspase-3 Rn00563902_m1 100±25.0 98.51±15.1 0.958 ITGAX Integrin subunit αX Rn01511082_m1 100±27.1 101.80±25.4 0.965 IL10 Interleukin 10 Rn01483988_g1 100±23.0 205.80±41.4 0.077 ITGB3 Integrin subunit β3 Rn00596601_m1 100±24.3 135.00±34.1 0.476 NFKB Nuclear factor κ light chain enhancer of activated B cells Rn01399572_m1 100±20.8 135.00±26.0 0.365 MPO Myeloperoxidase Rn01460205_m1 100±29.5 1554.00±740.8 0.155 Open in a separate window Note: All day are normalized to ribosomal gene RPL32 and expressed as relative units.

Techniques:

Journal: The Journal of Neuroscience

Article Title: The Autism Protein Ube3A/E6AP Remodels Neuronal Dendritic Arborization via Caspase-Dependent Microtubule Destabilization

doi: 10.1523/JNEUROSCI.1511-17.2017

Figure Lengend Snippet: E6AP autism mouse model neurons show impairment in spine maturation and reduction in dendritic branching. A, Brain lysates collected from WT or 2X Tg mice at P15 were probed for E6AP, XIAP, caspase-3, cleaved caspase-3, cleaved tubulin, and total tubulin. GAPDH was also probed as a loading control. B–D, Quantification analysis of Western blots for XIAP, cleaved caspase-3, and cleaved tubulin; n = 3 for each. E, At P15, brains of WT and 2X Tg mice were subjected to Golgi staining. Representative images of spine morphology of layer-V somatosensory cortical neurons are shown. F, Mean spine density decreased in 2X Tg mice; n = 10 neurons. G, Mean spine length increased in 2X Tg mice; n = 10 neurons. H, I, The percentage and number of filopodia increased in 2X Tg mice; n = 10 neurons. J, Representative layer-V pyramidal neuron tracing images of Golgi staining from P15 WT and 2X Tg mouse brain slices. K, L, Measurement of average dendrite number and total dendritic length in pyramidal neurons; n = 12 neurons. Error bars represent SEM, *p < 0.05, **p < 0.01 (Fig. 7-1, ). Summary of the E6AP-dependent dendritic remodeling pathway.

Article Snippet: The following cDNA plasmids were obtained from Addgene: p4054-E6AP (#8658), E6AP C820A (#37602), pEBB-XIAP (#11558), and pCDNA3-caspase-3 C163A (#11814). mCherry-tubulin wild-type (WT) and mCherry-tubulin K40A were kind gifts from Dr. Saudou Frederic (Institut Curie). pHR-pTRE-iCre-mCherry and pHR-rtTA (Tet-ON) were generously provided by Wilson Wong.

Techniques: Western Blot, Staining

Journal: The Journal of Neuroscience

Article Title: The Autism Protein Ube3A/E6AP Remodels Neuronal Dendritic Arborization via Caspase-Dependent Microtubule Destabilization

doi: 10.1523/JNEUROSCI.1511-17.2017

Figure Lengend Snippet: Activation of caspase-3 is required for E6AP-dependent dendritic remodeling. A, Neurons were transfected with surfGFP (green; Control) or together with E6AP, and the cleaved (activated) caspase-3 (red) was immunostained 24 h later. Scale bar, 50 μm. B, Quantification of the cleaved caspase-3 immunofluorescence signals; n = 10. E6AP expression resulted in higher levels of cleaved caspase-3. C, D, DIV 2 hippocampal neurons were infected with AAV9 GFP virus or AAV9 E6AP virus for 10 d and cleaved caspase-3 levels were measured by Western blot. Neurons infected with E6AP virus showed higher levels of cleaved caspase-3; n = 3 independent experiments. E, Dendritic arborization reduction in E6AP neurons was blocked by inhibiting caspase-3 cleavage with the caspase-9 inhibitor Ac-LEHD-CMK (150 nm) at the time of transfection, as shown by Sholl analysis; n = 10. F, G, Neurons were transfected with surfGFP (control) or together with E6AP or E6AP + Casp3 C163A (E6AP + C3 mutant), a catalytic caspase-3 mutant. Scale bar, 50 μm. Sholl analysis revealed a rescue of the E6AP-induced dendritic remodeling by Casp3 C163A; n = 10. Error bars represent SEM, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Article Snippet: The following cDNA plasmids were obtained from Addgene: p4054-E6AP (#8658), E6AP C820A (#37602), pEBB-XIAP (#11558), and pCDNA3-caspase-3 C163A (#11814). mCherry-tubulin wild-type (WT) and mCherry-tubulin K40A were kind gifts from Dr. Saudou Frederic (Institut Curie). pHR-pTRE-iCre-mCherry and pHR-rtTA (Tet-ON) were generously provided by Wilson Wong.

Techniques: Activation Assay, Transfection, Immunofluorescence, Expressing, Infection, Western Blot, Mutagenesis

Journal: The Journal of Neuroscience

Article Title: The Autism Protein Ube3A/E6AP Remodels Neuronal Dendritic Arborization via Caspase-Dependent Microtubule Destabilization

doi: 10.1523/JNEUROSCI.1511-17.2017

Figure Lengend Snippet: Microtubule cleavage and retraction in E6AP-induced dendritic remodeling. A, Neurons were transfected with surfGFP and pTRE-E6AP for 24 h, and loaded with SiR-Tubulin, a fluorogenic and cell-permeable dye for tubulin labeling, before being treated with doxycycline (Dox) to induce E6AP expression. Tubulin and surfGFP images were obtained every 20 min for 12 h following Dox application. Representative images show that retraction of microtubule (red; hollow arrowhead) occurred before that of the GFP-positive dendritic branch (green; solid arrowhead). The original position of the dendritic tip is indicated by a dashed line. Scale bar, 5 μm. B, Neurons were infected with AAV9 GFP virus or AAV9 E6AP virus for 10 d, and cleaved tubulin levels were measured by Western blot with an antibody specifically against the cleaved microtubule (ΔTubulin). C, Quantification showed an increased level of microtubule cleavage in E6AP-infected neurons; n = 3 independent experiments. D, Representative image of E6AP neurons immunostained with ΔTubulin. Scale bar, 10 μm. E, Quantification of ΔTubulin immunointensity in neurons transfected with vector control, E6AP, or E6AP + Casp3 C163A (E6AP + C3 mutant), compared with control; n = 10. F, Morphology of neurons transfected with surfGFP, tubulin WT, E6AP, or E6AP + tubulin WT. Scale bar, 50 μm. G, Sholl analysis of dendritic arborization; n = 10 cells per condition. H, Quantification of total dendritic length; n = 10 cells per condition. Changes in tubulin stabilization also affected the E6AP-dependent dendritic remodeling (Fig. 5-1, . Error bars represent SEM, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Article Snippet: The following cDNA plasmids were obtained from Addgene: p4054-E6AP (#8658), E6AP C820A (#37602), pEBB-XIAP (#11558), and pCDNA3-caspase-3 C163A (#11814). mCherry-tubulin wild-type (WT) and mCherry-tubulin K40A were kind gifts from Dr. Saudou Frederic (Institut Curie). pHR-pTRE-iCre-mCherry and pHR-rtTA (Tet-ON) were generously provided by Wilson Wong.

Techniques: Transfection, Labeling, Expressing, Infection, Western Blot, Plasmid Preparation, Mutagenesis

Journal: Pakistan Journal of Zoology

Article Title: Mechanism of Antipyretic Effect of Saposhnikoviae Radix Based on Network Pharmacology and Experimental Verification

doi: 10.17582/journal.pjz/20220702200702

Figure Lengend Snippet: Fig. 6. The expression levels of COX2, TNF-α, BAX, caspase3 and BCL2 proteins in the hypothalamus of rats in each group. For details of groups, please refer to Table III.

Article Snippet: Lipopolysaccharide (Beijing Solebao Technology Co., Ltd., batch number: L8880), wild SR (provided by Baishan Vocational and Technical College, identified by Chinese Medicine Resources Teaching and Research Section of Changchun University of Traditional Chinese Medicine, batch number: 2020901), pentobarbital sodium (Guangzhou Chemical Reagent Factory, batch number: 850601), IL-1β, IL-6, TNF-α(ELISA MM-0190R1, MM-0180R1), COX2 antibody, caspase3 antibody, Bax antibody, BCL2 antibody, secondary antibody (Wuhan PROTEINTECH Biotechnology Co., Ltd., article number: 12375-1-AP, 66470-2-lg, 50599-2-lg, 26593-1) Article number: WL01581), RIPA lysate, BCA protein concentration determination kit (Shanghai Beyotime Biotechnology Co., Ltd., article number: P0013B, P0010), protease inhibitor mixture (Shanghai Yami Biomedical Technology Co., Ltd., article number: GRF101), PMSF (Wuhan Service Biotechnology Co., Ltd., article number: G2008), Freeze centrifuge (Hunan Xiangyi Laboratory Instrument Development Co., Ltd., model: TGL-16), Invitrogen iBright imaging system (Thermo Company, USA, model: CL750), PowerPacTMbasic electrophoresis apparatus (Bio-Rad Company, USA, model: 1645050).

Techniques: Expressing