Journal: Journal of Virology

Article Title: Extended Synaptotagmin 1 Interacts with Herpes Simplex Virus 1 Glycoprotein M and Negatively Modulates Virus-Induced Membrane Fusion

doi: 10.1128/jvi.01281-17

Figure Lengend Snippet: Fig. 3: E-Syt1 interacts with HSV-1gM in infected cells. HeLa cells were infected 728

Article Snippet: Primary antibodies and dilution used in this study were as follows: anti-336 myc rabbit polyclonal (1:1000; 2272, Cell Signaling Technology), anti-myc mouse 337 polyclonal ( 9B11) (1:100; 2276, Cell Signaling Technology) anti-HA mouse monoclonal 338 (1:1000; SC-7392, Santa Cruz), anti E-Syt1 rabbit polyclonal (1:1000; A303-362A, 339 Bethyl Laboratories), anti E-Syt2 rabbit polyclonal (1:1000; NBP1-59988, Novus 340 Biologicals), anti E-Syt3 rabbit polyclonal (1:1000; NBP1-91354, Novus Biologicals), 341 anti β-actin mouse monoclonal (1:2500; ab6276, Abcam), anti γ-Tubulin mouse 342 monoclonal (1:5000; T6557, Sigma-Aldrich), anti-HSV VP5 mouse monoclonal (1:2000; 343 Virusys), anti-HSV gM rabbit polyclonal PAS980 (1:1000; courtesy of Dr. Lynn 344 Enquist), anti-HSV gM rabbit polyclonal 4c10 (1:000; courtesy of Dr. Joel Baines) and 345 anti-human DDX3 rabbit R648 polyclonal (1:4000; courtesy of Dr. A. Patel).

Techniques: Infection

Journal: Journal of Virology

Article Title: Extended Synaptotagmin 1 Interacts with Herpes Simplex Virus 1 Glycoprotein M and Negatively Modulates Virus-Induced Membrane Fusion

doi: 10.1128/jvi.01281-17

Figure Lengend Snippet: Fig. 5: Impact of the virus on E-Syt1 expression. HeLa cells were mock treated or 750

Article Snippet: Primary antibodies and dilution used in this study were as follows: anti-336 myc rabbit polyclonal (1:1000; 2272, Cell Signaling Technology), anti-myc mouse 337 polyclonal ( 9B11) (1:100; 2276, Cell Signaling Technology) anti-HA mouse monoclonal 338 (1:1000; SC-7392, Santa Cruz), anti E-Syt1 rabbit polyclonal (1:1000; A303-362A, 339 Bethyl Laboratories), anti E-Syt2 rabbit polyclonal (1:1000; NBP1-59988, Novus 340 Biologicals), anti E-Syt3 rabbit polyclonal (1:1000; NBP1-91354, Novus Biologicals), 341 anti β-actin mouse monoclonal (1:2500; ab6276, Abcam), anti γ-Tubulin mouse 342 monoclonal (1:5000; T6557, Sigma-Aldrich), anti-HSV VP5 mouse monoclonal (1:2000; 343 Virusys), anti-HSV gM rabbit polyclonal PAS980 (1:1000; courtesy of Dr. Lynn 344 Enquist), anti-HSV gM rabbit polyclonal 4c10 (1:000; courtesy of Dr. Joel Baines) and 345 anti-human DDX3 rabbit R648 polyclonal (1:4000; courtesy of Dr. A. Patel).

Techniques: Virus, Expressing

Journal: Journal of Virology

Article Title: Extended Synaptotagmin 1 Interacts with Herpes Simplex Virus 1 Glycoprotein M and Negatively Modulates Virus-Induced Membrane Fusion

doi: 10.1128/jvi.01281-17

Figure Lengend Snippet: Fig. 9: E-Syt1 and E-Syt3 down regulation increases cell-to-cell spread. The 798

Article Snippet: Primary antibodies and dilution used in this study were as follows: anti-336 myc rabbit polyclonal (1:1000; 2272, Cell Signaling Technology), anti-myc mouse 337 polyclonal ( 9B11) (1:100; 2276, Cell Signaling Technology) anti-HA mouse monoclonal 338 (1:1000; SC-7392, Santa Cruz), anti E-Syt1 rabbit polyclonal (1:1000; A303-362A, 339 Bethyl Laboratories), anti E-Syt2 rabbit polyclonal (1:1000; NBP1-59988, Novus 340 Biologicals), anti E-Syt3 rabbit polyclonal (1:1000; NBP1-91354, Novus Biologicals), 341 anti β-actin mouse monoclonal (1:2500; ab6276, Abcam), anti γ-Tubulin mouse 342 monoclonal (1:5000; T6557, Sigma-Aldrich), anti-HSV VP5 mouse monoclonal (1:2000; 343 Virusys), anti-HSV gM rabbit polyclonal PAS980 (1:1000; courtesy of Dr. Lynn 344 Enquist), anti-HSV gM rabbit polyclonal 4c10 (1:000; courtesy of Dr. Joel Baines) and 345 anti-human DDX3 rabbit R648 polyclonal (1:4000; courtesy of Dr. A. Patel).

Techniques:

Journal: Journal of Virology

Article Title: Extended Synaptotagmin 1 Interacts with Herpes Simplex Virus 1 Glycoprotein M and Negatively Modulates Virus-Induced Membrane Fusion

doi: 10.1128/jvi.01281-17

Figure Lengend Snippet: Fig. 11: Overexpression of E-Syt1 or E-Syt-3 reduces viral yields. HeLa cells 818

Article Snippet: Primary antibodies and dilution used in this study were as follows: anti-336 myc rabbit polyclonal (1:1000; 2272, Cell Signaling Technology), anti-myc mouse 337 polyclonal ( 9B11) (1:100; 2276, Cell Signaling Technology) anti-HA mouse monoclonal 338 (1:1000; SC-7392, Santa Cruz), anti E-Syt1 rabbit polyclonal (1:1000; A303-362A, 339 Bethyl Laboratories), anti E-Syt2 rabbit polyclonal (1:1000; NBP1-59988, Novus 340 Biologicals), anti E-Syt3 rabbit polyclonal (1:1000; NBP1-91354, Novus Biologicals), 341 anti β-actin mouse monoclonal (1:2500; ab6276, Abcam), anti γ-Tubulin mouse 342 monoclonal (1:5000; T6557, Sigma-Aldrich), anti-HSV VP5 mouse monoclonal (1:2000; 343 Virusys), anti-HSV gM rabbit polyclonal PAS980 (1:1000; courtesy of Dr. Lynn 344 Enquist), anti-HSV gM rabbit polyclonal 4c10 (1:000; courtesy of Dr. Joel Baines) and 345 anti-human DDX3 rabbit R648 polyclonal (1:4000; courtesy of Dr. A. Patel).

Techniques: Over Expression

Journal: Journal of Virology

Article Title: Extended Synaptotagmin 1 Interacts with Herpes Simplex Virus 1 Glycoprotein M and Negatively Modulates Virus-Induced Membrane Fusion

doi: 10.1128/jvi.01281-17

Figure Lengend Snippet: Fig. 13: E-Syt1 and E-Syt3 knockdown increase cell-to-cell fusion in infected 843

Article Snippet: Primary antibodies and dilution used in this study were as follows: anti-336 myc rabbit polyclonal (1:1000; 2272, Cell Signaling Technology), anti-myc mouse 337 polyclonal ( 9B11) (1:100; 2276, Cell Signaling Technology) anti-HA mouse monoclonal 338 (1:1000; SC-7392, Santa Cruz), anti E-Syt1 rabbit polyclonal (1:1000; A303-362A, 339 Bethyl Laboratories), anti E-Syt2 rabbit polyclonal (1:1000; NBP1-59988, Novus 340 Biologicals), anti E-Syt3 rabbit polyclonal (1:1000; NBP1-91354, Novus Biologicals), 341 anti β-actin mouse monoclonal (1:2500; ab6276, Abcam), anti γ-Tubulin mouse 342 monoclonal (1:5000; T6557, Sigma-Aldrich), anti-HSV VP5 mouse monoclonal (1:2000; 343 Virusys), anti-HSV gM rabbit polyclonal PAS980 (1:1000; courtesy of Dr. Lynn 344 Enquist), anti-HSV gM rabbit polyclonal 4c10 (1:000; courtesy of Dr. Joel Baines) and 345 anti-human DDX3 rabbit R648 polyclonal (1:4000; courtesy of Dr. A. Patel).

Techniques: Knockdown, Infection

Journal: The Journal of Cell Biology

Article Title: The intracellular translocation of the components of the fibroblast growth factor 1 release complex precedes their assembly prior to export

doi: 10.1083/jcb.200203084

Figure Lengend Snippet: Dominant-negative mutants of S100A13 and Syt1 do not affect the heat shock–induced redistribution of cytosolic FGF1:GFP. (A) NIH 3T3 cells were transiently cotransfected with FGF1:GFP and either S100A13:Myc (a–f) or S100A13Δ88–98:Myc (g–l), and 48 h later the cells were fixed after a 2-h incubation at either 37 (a–c and g–i) or 42°C (d–f and j–l). The cells were stained with an anti-Myc antibody followed by an Alexa-647–conjugated secondary antibody. Confocal images of median horizontal cell sections were taken using the 100X objective and the intracellular distribution of FGF1:GFP (a, d, g, and j), S100A13:Myc (b and e), S100A13Δ88–98Myc (h and k), and their respective overlays (c, f, i, and l) are shown. Bar, 10 μM. (B) NIH 3T3 cells were transiently cotransfected with FGF1:GFP and either Syt1:Myc (a–f) or Syt1Δ120–214:Myc (g–l) and 48 h later the cells were fixed after 2 h of incubation at either 37 (a–c and g–i) or 42°C (d–f and j–l). The cells were stained with either an anti-Syt1 (b and e) or an anti-Myc (h and k) antibody followed by an Alexa 647–conjugated secondary antibody. Confocal images of median horizontal cell sections were taken using the 100× objective and the intracellular distribution of FGF1:GFP (a, d, g, and j), Syt1:Myc (b and e), Syt1Δ120–214:Myc (h and k), and their respective overlays (c, f, i, and l) are shown. Bar, 10 μM.

Article Snippet: A monoclonal antibody to the extravesicular fragment of Syt1 (Synaptic Systems) followed with an Alexa 647–conjugated anti–mouse IgG antibody was used to detect wild-type Syt1 and rabbit antibodies against Erk1 and Erk2 were obtained from Santa Cruz Biotechnology, Inc.

Techniques: Dominant Negative Mutation, Incubation, Staining

Journal: The Journal of Headache and Pain

Article Title: Repeated oxytocin prevents central sensitization by regulating synaptic plasticity via oxytocin receptor in a chronic migraine mouse model

doi: 10.1186/s10194-021-01299-3

Figure Lengend Snippet: Effect of OT on synapse-associated protein expression levels. A Representative WB bands showing the expression of PSD-95, syt-1, and snap25. B , C , and D Quantification of PSD-95 ( B ), syt-1 ( C ), and snap25 ( D ) protein levels showed that these proteins were significantly increased in the NTG + vehicle group compared to the sham group. The expression levels of these three proteins after OT treatment were reduced, while L368,899 blocked these effects. One-way ANOVA with Dunnett’s post hoc test, n = 6/group; *** p < 0.001 vs. the sham group; ### p < 0.001 vs. the NTG + vehicle group; && p < 0.01, &&& p < 0.001 vs. the NTG + OT group

Article Snippet: The membranes were blocked for 2 h at room temperature in Tris-buffered saline with Tween-20 (TBST) containing 5 % nonfat milk and incubated overnight at 4 °C with the following primary antibodies: anti-OTR (1:500; Abcam, Cambridge, UK), anti-calcitonin gene-related peptide (CGRP; 1:3000; Abcam, Cambridge, UK), anti-c-fos (1:1000; Abcam, Cambridge, UK), anti-PSD-95 (1:1000; Cell Signaling Technology, Massachusetts, USA), anti-syt-1 (1:1000; Bioss, Beijing, China), anti-snap25 (1:4000; Abcam, Cambridge, UK), anti-NR2B (1:1000; Proteintech, Illinois, USA), anti-NR2B phosphor (pNR2B-Y1472; 1:500; Bioss, Beijing, China), anti-NR2B phosphor (pNR2B-Y1252; 1:1000; Abcam Cambridge, UK), anti-AC1 (1:500; Abcam, Cambridge, UK), anti-PKA C-α (1:3000; Cell Signaling Technology, Massachusetts, USA), anti-CREB (1:1000; Wanlei Bio, Shenyang, China), anti-CREB phosphor (pCREB ; 1:5000; Abcam, Cambridge, UK), and anti-Glyceraldehyde-3-phosphate dehydrogenase (GAPDH; 1:5000; Zen-Bioscience, Chengdu, China).

Techniques: Expressing

Journal: Frontiers in Behavioral Neuroscience

Article Title: Altered cognition and anxiety in adolescent offspring whose mothers underwent different-pattern maternal sleep deprivation, and cognition link to hippocampal expressions of Bdnf and Syt-1

doi: 10.3389/fnbeh.2022.1066725

Figure Lengend Snippet: Effects of MSD on the expression of hippocampal of BDNF and Syt-1 proteins in the offspring. (A) The expression levels of BDNF ( n = 12, 6 females and 6 males in each treatment group) and Syt-1 ( n = 12, 6 females and 6 males in each treatment group) proteins in the hippocampus at 2 months. The results of protein quantification are depicted in (B,C) . ** P < 0.01 vs. Male CON group; ## P < 0.01 vs. Male CSD3h group; $$ P < 0.01 vs. Male CSD6h group; aa P < 0.01 vs. Female CON group; bb P < 0.01 vs. Female CSD3h group; cc P < 0.01 vs. Female CSD6h group.

Article Snippet: The membrane was subsequently incubated with the rabbit anti-BDNF (1:1000; Abcam, Cambridge, UK) and rabbit anti-Syt-1 (1:1000; Bioss, Beijing, China) primary antibodies, and the horseradish peroxidase (HRP) labeled goat anti-rabbit IgG (Zsbio, ZB-2301) secondary antibody, according to the manufacturer's instructions.

Techniques: Expressing

Journal: Frontiers in Behavioral Neuroscience

Article Title: Altered cognition and anxiety in adolescent offspring whose mothers underwent different-pattern maternal sleep deprivation, and cognition link to hippocampal expressions of Bdnf and Syt-1

doi: 10.3389/fnbeh.2022.1066725

Figure Lengend Snippet: The effects of MSD on the expression of hippocampal of BDNF and Syt-1 mRNA in the offspring. The mRNA expression levels of BDNF ( n = 16, 8 females and 8 males in each treatment group) and Syt1 ( n = 16, 8 females and 8 males in each treatment group) in the hippocampus at 2 months. The results of mRNA quantification are depicted in (A,B) . ** P < 0.01 vs. Male CON group; # P < 0.05 and ## P < 0.01 vs. Male CSD3h group; $ P < 0.05 and $$ P < 0.01 vs. Male CSD6h group; a P < 0.05 and aa P < 0.01 vs. Female CON group; b P < 0.05 and bb P < 0.01 vs. Female CSD3h group; c P < 0.05 and cc P < 0.01 vs. Female CSD6h group.

Article Snippet: The membrane was subsequently incubated with the rabbit anti-BDNF (1:1000; Abcam, Cambridge, UK) and rabbit anti-Syt-1 (1:1000; Bioss, Beijing, China) primary antibodies, and the horseradish peroxidase (HRP) labeled goat anti-rabbit IgG (Zsbio, ZB-2301) secondary antibody, according to the manufacturer's instructions.

Techniques: Expressing

Journal: Molecular Neurobiology

Article Title: Pathological Deficit of Cystatin B Impairs Synaptic Plasticity in EPM1 Human Cerebral Organoids

doi: 10.1007/s12035-023-03812-y

Figure Lengend Snippet: Synaptic proteins in human cerebral organoids and neurons. a Western blots of the total lysate (h) and synaptosomes (syn) from hCOs using synaptophysin (SYP), sintaxin (STX), synaptotagmin 1/2 (SYT), and β-actin (ACTB) antibodies. Molecular weight in kDalton (kDa) is indicated on the left. Enrichment of b synaptophysin, c syntaxin, and d synaptotagmin 1/2 protein level, normalized on β-actin, in the synaptosomes (syn) compared to the homogenate (h, dotted line) of hCOs at 2, 3, 5, 7, 11, and 14 months (m). e Western blot of the synaptosomes from hCOs using cystatin B (CSTB) and ACTB antibodies. Molecular weight in kDa is indicated on the left. f CSTB protein level, normalized on β-actin, in synaptosomes of hCOs. m, months. Each time point is a pool of 20–40 hCOs. g Micrographs showing doublecortin (DCX, red), cystatin B (CSTB, white), and synaptophysin (SYP, green) in 10-week neurons from NPCs. DAPI (blu) stained nuclei. The arrowheads in the enlarged images of the boxed area indicate the colocalization of SYP and CSTB. Scale bar in each panel

Article Snippet: Western blot analysis was performed as previously reported [ , , ] with the following primary antibodies: SYP (1:1000, AB9272 Millipore), syntaxin (STX, 1:500, E-AB-33012 Elabscience), synaptotagmin 1/2 (SYT, 1:1000, E-AB-33005 Elabscience), CSTB (1:2000, Antikoerper AbIN271833), CD81 (1:500, AB9272 Santa Cruz), CD9 (1:500, sc-13118 Santa Cruz Biotechnology), CD82 (1:500, sc-518002 Santa Cruz Biotechnology), eIF4G2 (1:1000, HPA016965 Sigma-Aldrich), and β-actin (ACTB, 1:2000, 612,656 BD Biosciences). β-actin was used as a normalizer since its expression levels showed no substantial variations among the control and patients’ hCOs in both lysate and synaptosomal fraction (data not shown).

Techniques: Western Blot, Molecular Weight, Staining

Journal: Molecular Neurobiology

Article Title: Pathological Deficit of Cystatin B Impairs Synaptic Plasticity in EPM1 Human Cerebral Organoids

doi: 10.1007/s12035-023-03812-y

Figure Lengend Snippet: Pathological low expression levels of CSTB results in synaptic impairment in EPM1 hCOs. Western blots of a total lysate and b synaptosomal fractions from controls and EPM1 hCOs at different developmental stages using cystatin B (CSTB) and β-actin (ACTB) antibodies. Western blots of c total lysate and d synaptosomal fractions from controls and EPM1 hCOs using eukariotic Initiation Factor 4G2 (eIF4G2) and ACTB antibodies. Western blots (upper panels) and relative quantifications (lower panels). e Western blots of synaptosomes from controls and EPM1 hCOs at different developmental stages, using synaptophysin (SYP), sintaxin (STX), synaptotagmin 1/2 (SYT), and ACTB antibodies and relative quantifications. Molecular weight of each protein is indicated on the left (kDa). Data are presented as mean ± SD from n = 2 pools of 20–40 organoids. Controls: C1, C2; patients: E1, E2; m, months

Article Snippet: Western blot analysis was performed as previously reported [ , , ] with the following primary antibodies: SYP (1:1000, AB9272 Millipore), syntaxin (STX, 1:500, E-AB-33012 Elabscience), synaptotagmin 1/2 (SYT, 1:1000, E-AB-33005 Elabscience), CSTB (1:2000, Antikoerper AbIN271833), CD81 (1:500, AB9272 Santa Cruz), CD9 (1:500, sc-13118 Santa Cruz Biotechnology), CD82 (1:500, sc-518002 Santa Cruz Biotechnology), eIF4G2 (1:1000, HPA016965 Sigma-Aldrich), and β-actin (ACTB, 1:2000, 612,656 BD Biosciences). β-actin was used as a normalizer since its expression levels showed no substantial variations among the control and patients’ hCOs in both lysate and synaptosomal fraction (data not shown).

Techniques: Expressing, Western Blot, Molecular Weight