Journal: Journal of Nanophotonics

Article Title: Mouse neuroblastoma cell-based model and the effect of epileptic events on calcium oscillations and neural spikes

doi: 10.1117/1.jnp.7.073794

Figure Lengend Snippet: Fig. 2 Fluorescent imaging of mouse neuroblastoma neuro2a (N2A) cells using fluo-4 NW: (a) control cells; (b) 30 min after the injection of 4-AP (10 mM); and (c) 30 min after the injection of 4-AP (50 mM).

Article Snippet: 2.1 Mouse Neuroblastoma N2A Cell Culture The mouse neuroblastoma N2A cells were purchased from American Type Culture Collection (CCl-131, Manassas, Virginia) and cultured at a density of 1.0 × 104 in Dulbecco’s modified Eagle’s medium (DMEM) that was supplemented with heat-inactivated 10% fetal bovine Journal of Nanophotonics 073794-2 Vol.

Techniques: Imaging, Control, Injection

Journal: Journal of Nanophotonics

Article Title: Mouse neuroblastoma cell-based model and the effect of epileptic events on calcium oscillations and neural spikes

doi: 10.1117/1.jnp.7.073794

Figure Lengend Snippet: Fig. 3 Calcium oscillations in mouse neuroblastoma N2A cells: control cells (blue line), 30 min after the injection of the 4-AP with 10 mM (black line) and 50 mM (red line).

Article Snippet: 2.1 Mouse Neuroblastoma N2A Cell Culture The mouse neuroblastoma N2A cells were purchased from American Type Culture Collection (CCl-131, Manassas, Virginia) and cultured at a density of 1.0 × 104 in Dulbecco’s modified Eagle’s medium (DMEM) that was supplemented with heat-inactivated 10% fetal bovine Journal of Nanophotonics 073794-2 Vol.

Techniques: Control, Injection

Journal: The Journal of Neuroscience

Article Title: A Multilevel Functional Study of a SNAP25 At-Risk Variant for Bipolar Disorder and Schizophrenia

doi: 10.1523/JNEUROSCI.1040-17.2017

Figure Lengend Snippet: In vitro analysis of rs6039769 in N2a cells. Quantification of the SNAP25 expression level was estimated using a luciferase reporter gene under control of the human SNAP25 promoter region containing either the C or the A allele of rs6039769. Both constructs showed a higher reporter expression level than the empty vector. The C allele showed a 20% increase of the reporter expression level than the A allele. The values are reported as the mean ± SD. ***p < 0.0001.

Article Snippet: Functional in vitro study The Neuro-2a (N2a) mouse neuroblastoma cell line (catalog #400394/p451_Neuro-2A, CLS; RRID: CVCL_0470 ) was grown at 37°C, with 5% CO 2 in DMEM containing GlutaMAX (Thermo Fisher Scientific) and was supplemented with 10% fetal bovine serum, 100 U/ml penicillin, and 100 μg/ml streptomycin.

Techniques: In Vitro, Expressing, Luciferase, Control, Construct, Plasmid Preparation

Journal: The Journal of Clinical Investigation

Article Title: Protein kinase N1 critically regulates cerebellar development and long-term function

doi: 10.1172/JCI96165

Figure Lengend Snippet: ( A ) Phospho-AKT (p-AKT) [T308] intensity was measured in untransfected Pkn1 –/– Cgcs and Pkn1 –/– Cgcs expressing human HA-tagged PKN1 (hPKN1) [8–22 transfected cells were analyzed per experiment; 2-tailed paired t test, t = 5.365, * P = 0.033, n = 3 from 3 litters]. ( B ) Cgc axonal length was measured at DIV1 after 24 hours of treatment with the AKT inhibitor MK-2206 (1 or 5 μM) in TAU-stained Cgcs [1-way ANOVA with Newman-Keuls multiple-comparisons test, F (5,15) = 26.97, P < 0.0001, post-test ** P < 0.01, *** P < 0.001; n = 3–4 WT, 3–4 Pkn1 –/– Cgc preparations from 3–4 litters per group]. ( C ) NeuroD2 expression levels were analyzed in Cgcs at DIV6–8 [2-tailed unpaired t test, t = 3.228, ** P = 0.008, n = 7 WT, 6 Pkn1 –/– Cgc preparations from 6–7 different litters]. Representative WIS-NeuroMath–analyzed output images are shown. ( D ) NeuroD2 intensity was measured at DIV4 in untransfected Pkn1 –/– Cgcs and Pkn1 –/– Cgcs expressing hPKN1 [26–32 transfected cells were analyzed per experiment; 2-tailed paired t test, t = 4.904, * P = 0.0392, n = 3 from 3 litters]. ( E ) WT Cgc protein extracts at DIV1 were analyzed for the effect of 24 hours of treatment with MK-2206 on NeuroD2 protein levels [1-way ANOVA with Newman-Keuls multiple-comparisons test, F (2,8) = 11.76, P = 0.0042, post-test ** P < 0.01, n = 3–4 from 3–4 litters]. All data are presented as individual n values with mean ± SEM. Scale bars: 10 μm in A and D , 50 μm in B . Experimenters were not blinded to the genotype or treatment.

Article Snippet: VGlut1 (catalog 48-2400), goat anti-rabbit–Alexa Fluor 488 (catalog A11070), and goat anti-mouse–Alexa Fluor 555 (catalog A21425) were purchased from Thermo Fisher Scientific, and GluD2 (D-9, catalog sc-393437) and NeuroD2 (G-10, catalog sc-365896) were from Santa Cruz Biotechnology.

Techniques: Expressing, Transfection, Staining

Journal: The Journal of Clinical Investigation

Article Title: Protein kinase N1 critically regulates cerebellar development and long-term function

doi: 10.1172/JCI96165

Figure Lengend Snippet: ( A ) siRNAs targeting Akt123 reduce pan-AKT expression after DIV1. Pictures are representative of 5 separate experiments. For analysis at DIV1 and DIV4 in WT and Pkn1 –/– Cgcs, see . ( B ) siRNAs targeting Akt123 significantly reduce axonal length of Pkn1 –/– Cgcs at DIV1 [1-way ANOVA with Newman-Keuls multiple-comparisons test, F (3,16) = 20.78, P < 0.0001, post-test *** P < 0.001, n = 5 WT, 5 Pkn1 –/– Cgc preparations from 3–5 litters per group]. Axons were stained with TAU. Representative WIS-NeuroMath–analyzed output images are shown. ( C ) siRNAs targeting Akt123 significantly reduce NeuroD2 intensity in WT and Pkn1 –/– Cgcs at DIV4 [1-way ANOVA with Newman-Keuls multiple-comparisons test, F (3,16) = 18.73, P < 0.0001, post-test * P < 0.05, *** P < 0.001; n = 5 WT, 5 Pkn1 –/– Cgc preparations from 3–5 litters per group]. ( D ) Presynaptic sites (varicosities) were stained with TAU and synapsin I. Insets represent higher-magnification single- and double-labeled examples of axonal varicosities (indicated by arrows). White varicosities in double-labeled insets demonstrate TAU and synapsin I colocalization. siRNAs targeting Akt123 significantly increase the density of presynaptic sites in Pkn1 –/– Cgcs at DIV4 [1-way ANOVA with Newman-Keuls multiple-comparisons test, F (3,16) = 16.62, P < 0.0001, post-test *** P < 0.001, n = 5 WT, 5 Pkn1 –/– Cgc preparations from 3–5 litters per group]. All data are presented as individual n values with mean ± SEM. Scale bars: 50 μm, except in inset in D : 10 μm. Experimenters were not blinded to the genotype or treatment.

Article Snippet: VGlut1 (catalog 48-2400), goat anti-rabbit–Alexa Fluor 488 (catalog A11070), and goat anti-mouse–Alexa Fluor 555 (catalog A21425) were purchased from Thermo Fisher Scientific, and GluD2 (D-9, catalog sc-393437) and NeuroD2 (G-10, catalog sc-365896) were from Santa Cruz Biotechnology.

Techniques: Expressing, Staining, Labeling

Journal: The Journal of Clinical Investigation

Article Title: Protein kinase N1 critically regulates cerebellar development and long-term function

doi: 10.1172/JCI96165

Figure Lengend Snippet: ( A ) There was a significant correlation between the p-AKT [S473]/AKT ratio and the PKN1/GAPDH ratio in cerebellar protein extracts from P1–P15 WT animals (Pearson correlation, number of XY pairs: 4, Pearson r = –0.9990, 2-tailed, P = 0.001, R 2 = 0.9979, n = 3–4 from 3–4 litters per group). Data are presented as mean ± SEM. ( B and C ) Confocal images of cerebellar sections of P8 and P15 WT and Pkn1 –/– animals stained for p-AKT [S473] and Hoechst (P8) ( B ) or p-AKT [S473] and VGlut2 (P15) ( C ). Pictures are representative of at least 3 independent experiments. ( D ) Western blot analysis of NeuroD2 expression in P8 [2-tailed unpaired t test, t = 2.546, * P = 0.0314, n = 5 WT, 6 Pkn1 –/– animals from 4–5 different litters] and in P15 WT and Pkn1 –/– whole cerebella protein extracts [2-tailed unpaired t test, t = 2.541, ** P = 0.0440, n = 4 WT, 4 Pkn1 –/– animals from 4–5 different litters]. Data are presented as individual n values with mean ± SEM. All scale bars: 50 μm. Experimenters were not blinded to the genotype.

Article Snippet: VGlut1 (catalog 48-2400), goat anti-rabbit–Alexa Fluor 488 (catalog A11070), and goat anti-mouse–Alexa Fluor 555 (catalog A21425) were purchased from Thermo Fisher Scientific, and GluD2 (D-9, catalog sc-393437) and NeuroD2 (G-10, catalog sc-365896) were from Santa Cruz Biotechnology.

Techniques: Staining, Western Blot, Expressing