Journal: The Journal of Neuroscience

Article Title: Brain-Derived Neurotrophic Factor Promotes the Maturation of GABAergic Mechanisms in Cultured Hippocampal Neurons

doi: 10.1523/JNEUROSCI.22-17-07580.2002

Figure Lengend Snippet: BDNF upregulates GABAA receptor (GABAaR) protein and presynaptic proteins. Cells were cultured in the presence or absence of 50 ng/ml BDNF for 7–10 d, and the extracts were analyzed by Western blot for NR1, GluR1, and GABAAR β2/3 (A), and syntaxin, synaptobrevin, and synaptophysin (B). Immunoreactive signals were normalized to the control. Insets are representative Western blots. A blot for β-actin for normalization is also shown. Data are means ± SEM of three independent experiments. *p < 0.05 versus control, Student's t test.

Article Snippet: The membrane was incubated in PBS containing 0.05% Tween 20 and 5% nonfat dry milk for 1 hr and then exposed to a primary antibody against synaptophysin (1:200, mouse; Boehringer Mannheim Biochemica), synaptobrevin (1:500, rabbit; Wako, Osaka, Japan), syntaxin (1:300, mouse; Wako), GluR1 (1:300, rabbit;Chemicon), NR1 (1:300, rabbit; Chemicon), or GABA A Rβ2/3 (only the β3 subunit was detected in our culture; 1:300, mouse; Upstate Biotechnology, New York, NY) in 0.05% Tween 20–PBS at 4°C overnight and then to horseradish peroxidase–conjugated secondary antibody for appropriate IgG.

Techniques: Cell Culture, Western Blot

Journal: Nature Communications

Article Title: Functional exploration of colorectal cancer genomes using Drosophila

doi: 10.1038/ncomms13615

Figure Lengend Snippet: ( a , b ) Plot of P -values indicating significance of compound rescue ( a ) and summary of compound response ( b ) of ras G12V and ras G12V p53 Ri pten Ri apc Ri animals. P -values ( a ) were obtained by comparing the dissemination phenotype after compound feeding to dimethyl sulfoxide (DMSO) fed flies (dissemination plots can be found in ). Blue dots represent statistically significant results. ( c ) Quantification of dissemination in ras G12V pten Ri and ras G12V p53 Ri apc Ri animals treated with BEZ235. ( d ) Western blot analysis of PI3K pathway output from hindguts with indicated genotypes 7 days after induction of transgenes and quantification. Syn, Syntaxin (loading control). ( e ) Time-course analysis of PI3K pathway activation status in control and ras G12V p53 Ri pten Ri apc Ri hindguts. ( f ) Western blot analysis of the biochemical response by ras G12V and ras G12V p53 Ri pten Ri apc Ri animals to PI3K pathway inhibitors. ( g ) Quantification of dissemination in indicated genotypes treated with BEZ235 or DMSO. ( h ) Schematic illustration of the mechanism of resistance to BEZ235: genetically activating mTORC1 promotes BEZ235 sensitivity. ( d , e ) Each data point represents the average response of two to five biological replicates with ten hindguts per replicate; error bars: s.e.m. ( a – c , g ) n =2 replicates, 30 flies per replicate; error bars: s.e.m. * P <0.01 and ** P <0.05 (Fisher's exact test). Compound doses reflect concentrations in the food. Uncropped gels with molecular markers for d and f can be found in .

Article Snippet: Primary antibodies were as follows: rabbit anti Drosophila phospho-AKT (p-Ser505; Drosophila equivalent of mammalian AKT p-Ser473, 1:1,000, Cell Signaling, catalogue number 4054S), rabbit anti-mouse AKT (1:1,000, Cell Signaling, catalogue number 4691S), rabbit anti-mouse phospho-4EBP (Thr37/46, 1:1,000, Cell Signaling, catalogue number 2855S), rabbit anti-human phospho-AKT (Ser473, 1:1,000, Cell Signaling, catalogue number 4060S), mouse anti-chicken α-actin (1:1,000, DSHB catalogue number JLA20-s), rabbit anti-GFP (1:2,000, Sigma, catalogue number G1544), rabbit anti Drosophila P53 (1:1,000, DSHB, catalogue number H3), rabbit anti Drosophila Pten (1:1,000, gift from A. Wodarz ), mouse anti-dpERK (Thr183/Tyr185, 1:2,000, Sigma, catalogue number M8159), rabbit anti human phospho-S6 (Ser 235/236, 1:1,000, Cell Signaling, catalogue number 2211) and mouse anti Drosophila Syntaxin (1:1,000, DSHB, catalogue number 8C3).

Techniques: Western Blot, Control, Activation Assay

Journal: Nature Communications

Article Title: Functional exploration of colorectal cancer genomes using Drosophila

doi: 10.1038/ncomms13615

Figure Lengend Snippet: ( a ) Western blot analysis of PI3K signalling pathway output in ras G12V and ras G12V p53 Ri pten Ri apc Ri hindguts after 1 day feeding of SC79 at indicated doses. Syn, Syntaxin (loading control);ten hindguts per replicate. ( b ) Quantification of dissemination in ras G12V and ras G12V p53 Ri pten Ri apc Ri animals after sequential treatment with BEZ235 and indicated doses of SC79. ( c ) Quantification of dissemination in ras G12V and ras G12V p53 Ri pten Ri apc Ri animals after two different treatment schedules of SC79/BEZ235 and each drug alone. ( b , c ) n =2 replicates, 30 flies per replicate; error bars: s.e.m. * P <0.01 and ** P <0.05 (Fisher's exact test). ( b ) *Variable response; not all replicates show significant rescue. Drug doses reflect concentrations in the food. Uncropped gels used to generate panel a can be found in .

Article Snippet: Primary antibodies were as follows: rabbit anti Drosophila phospho-AKT (p-Ser505; Drosophila equivalent of mammalian AKT p-Ser473, 1:1,000, Cell Signaling, catalogue number 4054S), rabbit anti-mouse AKT (1:1,000, Cell Signaling, catalogue number 4691S), rabbit anti-mouse phospho-4EBP (Thr37/46, 1:1,000, Cell Signaling, catalogue number 2855S), rabbit anti-human phospho-AKT (Ser473, 1:1,000, Cell Signaling, catalogue number 4060S), mouse anti-chicken α-actin (1:1,000, DSHB catalogue number JLA20-s), rabbit anti-GFP (1:2,000, Sigma, catalogue number G1544), rabbit anti Drosophila P53 (1:1,000, DSHB, catalogue number H3), rabbit anti Drosophila Pten (1:1,000, gift from A. Wodarz ), mouse anti-dpERK (Thr183/Tyr185, 1:2,000, Sigma, catalogue number M8159), rabbit anti human phospho-S6 (Ser 235/236, 1:1,000, Cell Signaling, catalogue number 2211) and mouse anti Drosophila Syntaxin (1:1,000, DSHB, catalogue number 8C3).

Techniques: Western Blot, Control

Journal: Journal of neurochemistry

Article Title: Impaired AMPA receptor trafficking by a double knockout of zebrafish olfactomedin1a/b

doi: 10.1111/jnc.14231

Figure Lengend Snippet: Antibodies used in the study.

Article Snippet: AMPA and tricaine were purchased from Sigma. table ft1 table-wrap mode="anchored" t5 caption a7 Antibody to host Company Cat# Resource Identifier Used for (Work Dilution) Actin goat Santa Cruz sc-1616 RRID:AB_630836 WB (1:1,000) βIII tubulin chicken Abcam ab41489 RRID:AB_727049 WB (1:30,000) Flotillin1 mouse BD Transduction Lab. 610820 RRID:AB_398139 WB (1:250) Synaptophysin mouse Synaptic Systems 101011 RRID:AB_887824 WB (1:10,000) Syntaxin1 mouse Synaptic Systems 110011 RRID:AB_887844 WB (1:10,000), IP PSD95 mouse Thermo Fisher Sci.

Techniques: Transduction

Journal: Journal of neurochemistry

Article Title: Impaired AMPA receptor trafficking by a double knockout of zebrafish olfactomedin1a/b

doi: 10.1111/jnc.14231

Figure Lengend Snippet: Olfm1 interacts with presynaptic SNARE complex proteins and postsynaptic GluR2 in synaptosomes from adult wt brain. (a) Western blot analysis of indicated proteins in adult wt and olfm1 null zebrafish brain and retina. The blots marked by an asterisk was obtained in the P2 fraction. (b) Quantification of three independent experiments as in A for VAMP and synaptophysin. Data for each protein were normalized using beta-III tubulin level. **p < 0.01. (c) Biochemical fractionation of adult zebrafish brain and intracellular localization of olfm1 and some pre- and post-synaptic proteins in adult zebrafish brain. Note co-purification of olfm1 and several synaptosomal membrane proteins with the LP1 fraction but not with the soluble fraction (S3). (d) Co-immunoprecipitation of Olfm1 with GluR2 from the synaptosomal fraction. The levels of GluR2 and phosphorylated GluR2 were slightly increased in synaptosomes isolated from olfm1 null brain compared with wt brain. (e, f) Co-immunoprecipitation of Olfm1 with VAMP2 and syntaxin1 from brain synaptosomes.

Article Snippet: AMPA and tricaine were purchased from Sigma. table ft1 table-wrap mode="anchored" t5 caption a7 Antibody to host Company Cat# Resource Identifier Used for (Work Dilution) Actin goat Santa Cruz sc-1616 RRID:AB_630836 WB (1:1,000) βIII tubulin chicken Abcam ab41489 RRID:AB_727049 WB (1:30,000) Flotillin1 mouse BD Transduction Lab. 610820 RRID:AB_398139 WB (1:250) Synaptophysin mouse Synaptic Systems 101011 RRID:AB_887824 WB (1:10,000) Syntaxin1 mouse Synaptic Systems 110011 RRID:AB_887844 WB (1:10,000), IP PSD95 mouse Thermo Fisher Sci.

Techniques: Western Blot, Fractionation, Copurification, Immunoprecipitation, Isolation

Journal: International Journal of Molecular Sciences

Article Title: Prenatal Exposure to Valproic Acid Affects Microglia and Synaptic Ultrastructure in a Brain-Region-Specific Manner in Young-Adult Male Rats: Relevance to Autism Spectrum Disorders

doi: 10.3390/ijms21103576

Figure Lengend Snippet: Conditions used to perform the Western blot experiments.

Article Snippet: Mouse anti-Stx1 , Santa Cruz Biotechnology sc-12736 , 1:1000 5% milk in TBS-T 0.1%.

Techniques: Western Blot