Journal: Autophagy

Article Title: Table 1.

doi: 10.1080/15548627.2025.2597463

Figure Lengend Snippet: Autophagy-deficient PVALB-interneurons exhibit an altered synaptic composition, resulting in functional adaptations. (A) Representative images of surface GRIA2/GluA2 immunoreactivity (green) in hippocampal CA1 PVALB cells (magenta) from control ( Atg5 f/f ) vs. Pvalb-atg5 mice. Scale bar: 10 µm. (B) Quantification of the mean surface GRIA2/GluA2 intensity in control versus Pvalb-atg5 PVALB cells in hippocampal CA1. Results are expressed as a percentage of control values. Statistical analysis was performed using an unpaired, two-tailed Student’s t test (** p = 0.0028, t = 3.737, df = 12). Bars represent mean values ±SEM. N = 7 mice per genotype. (C) Representative images of total GRIA2/GluA2 immunoreactivity (green) in hippocampal CA1 PVALB cells (magenta) from control ( Atg5 f/f ) vs. Pvalb-atg5 mice. Scale bar: 10 µm. (D) Quantification of the mean total GRIA2/GluA2 intensity in control versus Pvalb-atg5 PVALB cells in hippocampal CA1. Results are expressed as a percentage of control values. Statistical analysis was performed using an unpaired, two-tailed Student’s t test ( ns p = 0.4154, t = 0.9076, df = 4). Bars represent mean values ±SEM. N = 3 mice per genotype. (E-I) Patch clamp recordings of AMPA (E) and NMDA (F) currents from CA1 PVALB cells from control ( Pvalb-TdT ) and Atg5 -deficient PVALB interneurons ( Pvalb-atg5-TdT ). These data were also plotted as the AMPA:NMDA ratio for controls and Pvalb-atg5-TdT animals, where all values were normalized to the control (G). (H) represents the average AMPA (black) and NMDA (gray) traces recorded at + 40 mV for control and Pvalb-atg5-TdT cells. (I) Reduced AMPA currents over NMDA currents in Pvalb-atg5-TdT mice when compared to controls. Black line indicates R 2 = 1. Statistical analyses were performed using a Student’s t test (for AMPA currents: * p = 0.0464, t = 2.100, df = 24; for NMDA currents, ns p = 0.3955, t = 0.8651, df = 24; for AMPA:NMDA ratios: * p = 0.0348, t = 2.238, df = 24). Bars represent mean values ± SEM. N = 3 animals per genotype ( Pvalb-TdT, n = 14; Pvalb-atg5-TdT , n = 16). (J) Current-clamp sample traces (400 pA injection, top) and the action potentials evoked by current steps (bottom) from PVALB neurons, from slices of control (Pvalb-TdT , RMP = −63 mV) and mutant Pvalb-atg5-TdT (RMP = −62 mV) mice. (K) Input-output plots from control (Pvalb-TdT) ( n = 14, N = 3) and Pvalb-atg5-TdT cells ( n = 16, N = 3). Statistical analysis was performed using a two-way repeated measures ANOVA followed by Sidak post hoc test; F(5,140) = 23.85 (**** p < 0.005). Bars represent mean values ± SEM. (L) Comparison of capacitance, resting membrane potential, action potential threshold and amplitude in control (Pvalb-TdT) ( n = 14, N = 3) and Pvalb-atg5-TdT cells ( n = 16, N = 3). Statistical analyses were performed using a Student’s t test (ap threshold, * p = 0.023, t = 2.398, df = 28). Bars represent mean values ± SEM. (M) Representative traces of sIPSC from slices of control (Pvalb-TdT) and mutant Pvalb-atg5-TdT mice recorded at + 5 mV. (N-O) Bar graphs and cumulative plots related tothe frequency and amplitude of sIpscs from control (Pvalb-TdT) ( n = 14, N = 3) and mutant Pvalb-atg5-TdT mice ( n = 16, N = 3). Statistical analysis was performed using an unpaired, two-tailed Student’s t test (* p = 0.0298, t = 2.289, df = 28 and ns p = 0.8654, t = 0.1710, df = 28, respectively). Bars represent mean values ± SEM. (P) Representative traces of paired-pulse ratio (IPSC 2 :IPSC 1 ) from three-month old control (Pvalb-TdT) and mutant Pvalb-atg5-TdT mice recorded at + 5 mV. (Q) Graph showing the paired-pulse ratio (IPSC 2 :IPSC 1 ) from control (Pvalb-TdT) ( n = 8, N = 2) and mutant Pvalb-atg5-TdT mice ( n = 12, N = 2). Statistical analysis was performed using an unpaired, two-tailed Student’s t test (* p = 0.0310, t = 2.340, df = 18). Bars represent mean values ± SEM. (R) Representative traces of excitation to inhibition ratio (EPSC −60 mV :IPSC +5 mV ) from control (Pvalb-TdT) and mutant Pvalb-atg5-TdT mice. (S) Graph showing the excitation to inhibition ratio (EPSC:IPSC) from control (Pvalb-TdT) ( n = 8, N = 2) and mutant Pvalb-atg5-TdT ( n = 14, N = 3). Statistical analysis was performed using an unpaired, two-tailed Student’s t test (* p = 0.0351, t = 2.261, df = 20). Bars represent mean values ± SEM.

Article Snippet: GRIA2/GluA2 (extracellular) , Guinea pig , Alomone labs , AGC-005-GP , 1:500 (IHC).

Techniques: Functional Assay, Control, Two Tailed Test, Patch Clamp, Injection, Mutagenesis, Comparison, Membrane, Inhibition

Journal: Nature

Article Title: Electrical and synaptic integration of glioma into neural circuits.

doi: 10.1038/s41586-019-1563-y

Figure Lengend Snippet: Fig. 4 | Glioma membrane depolarization promotes glioma progression. a, Optogenetic model for glioma depolarization. Blue dots represent ChR2-expressing glioma cells; light blue rectangle denotes region of analysis. P, postnatal day. b, Proliferation index of SU-DIPG- XIII-FL-ChR2 xenografts after mock stimulation or blue light stimulation, measured as percentage of GFP+/HNA+ cells expressing Ki67 (mock stim, n = 8; stim, n = 9 mice). c, As in b but for SU-DIPG-VI-ChR2 xenografts (n = 6 mice per group). d, Representative confocal micrographs from c, illustrating proliferation of SU-DIPG-VI-ChR2 xenografts. Red denotes human nuclei staining by HNA; white denotes Ki67. Scale bar, 50 µm. e, f, Kaplan–Meier survival curves of SU-DIPG-XIII-P* (P denotes pontine tumour) xenografts that overexpress GFP-only (green) or GluA2- WT-GFP (red) (e) and GFP-only (in 80% of cells, green) or GluA2-DN- GFP (in 80% of cells, blue) (f) (n = 5 mice per group). g, Competitive outgrowth of non-GluA2-DN-GFP-expressing cells in f, determined by

Article Snippet: We introduced SpeI (5′-TAAGCAACTAG TATGCAAAAGATTATGCAT-3′) and XmaI (5′-TGCTTACCCGGGC TAAATTTTAACACTTTCGAT-3′) restriction sites in full-length human GRIA2 clone (GluA2 WT) obtained from Origene (RC212599) by PCR.

Techniques: Membrane, Expressing, Staining

Journal: bioRxiv

Article Title: Single-molecule analysis reveals the rapid effect of estradiol on the surface movement of AMPAR in live neurons

doi: 10.1101/602813

Figure Lengend Snippet: Characterization of differentiated PC12 cells. ( a ) Photomicrograph depicting PC12 cells after 4 days of NGF treatment and ( b ) immunofluorescence staining of microtubule-associated protein 2 (MAP2) and β-III tubulin. Scale bars: 20 µm. ( c, d ) Representative recording of a whole-cell current-clamp step protocol obtained from PC12 neurons. ( c ) Application of increasing current steps in PC12 neuronal culture resulted in action potential generation. ( d ) Trace representing membrane potential changes in PC12 neurons. ( e, f ) Intensity profiles of a single ATTO 488-labeled GluR2-AMPAR and mGluR1 signal. The arrows indicate single-step photobleaching. ( g, h ) Histogram showing the intensity value of every spot found in a recording superimposed with a single fitted Gaussian curve (blue line).

Article Snippet: Before single-molecule imaging, PC12 neurons were incubated in dRPMI with ATTO 488-labeled antibodies directed against the extracellular N-terminal domain of rat GluR2 (1:100, Alomone Labs) or against the extracellular N-terminal domain of rat mGluR1 (1:100, Alomone Labs) at 37°C for 6 minutes.

Techniques: Immunofluorescence, Staining, Membrane, Labeling, Patch Clamp

Journal: bioRxiv

Article Title: Single-molecule analysis reveals the rapid effect of estradiol on the surface movement of AMPAR in live neurons

doi: 10.1101/602813

Figure Lengend Snippet: Single-molecule imaging of ATTO 488-labeled GluR2-AMPAR and mGluR1 receptors on neurites and on somas of PC12 neurons. ( a ) Representative trajectories of AMPAR molecules on somas and neurites. Scale bar: 2 µm. ( b ) The mean square displacement functions and trajectories represent AMPAR molecules with Brownian motion (red) and confined motion (blue). Scale bars: 0.1 µm. The cumulative probability functions of D values of AMPAR ( c ) and mGluR1 ( d ) on neurites and on somas are shown (n= 330-500 trajectories). **p<0.01, ***p<0.001

Article Snippet: Before single-molecule imaging, PC12 neurons were incubated in dRPMI with ATTO 488-labeled antibodies directed against the extracellular N-terminal domain of rat GluR2 (1:100, Alomone Labs) or against the extracellular N-terminal domain of rat mGluR1 (1:100, Alomone Labs) at 37°C for 6 minutes.

Techniques: Imaging, Labeling

Journal: bioRxiv

Article Title: Single-molecule analysis reveals the rapid effect of estradiol on the surface movement of AMPAR in live neurons

doi: 10.1101/602813

Figure Lengend Snippet: Effect of E2 on the surface trafficking of GluR2-AMPAR and mGluR1. The line graphs depict changes in the diffusion coefficient (D) of GluR2-AMPAR ( a ) and mGluR1 ( b ) molecules after vehicle or 100 pM E2 administration at different time points (mean ± SEM, n=100-128 trajectories per time point). ( c ) Distributions of D (as the percentage of vehicle control) of GluR2-AMPAR (average of 20-minute measurement) in the medium (dRPMI) or after administration of E2 at different concentrations (as the percent of the vehicle treatment expressed as the median ± 25-75%, n= 426-484 trajectories per group). ( d ) Distributions of D values of AMPAR on neurites in the presence of the estrogen receptor β (ERβ) agonist diarylpropionitrile (DPN), a GPER1 agonist (G1) and a GPER1 antagonist (G15) (expressed as the median ± 25-75%, n=427-468 trajectories). ( e ) Stochastic optical reconstruction microscopy images depicting immunolabeled AMPAR (magenta) and GPER1 (cyan) molecules on PC12 neurons. The dashed lines delineate the border between the neurite and soma. ( f ) Distributions of the minimal distance (greater than 500 nm) between GluR2-AMPAR and GPER1 after vehicle or E2 treatment (expressed as the median ± 25-75%, n=203-657). Scale bar: 2 µm; insert scale bar: 0.5 µm. **p<0.01, ***p<0.001

Article Snippet: Before single-molecule imaging, PC12 neurons were incubated in dRPMI with ATTO 488-labeled antibodies directed against the extracellular N-terminal domain of rat GluR2 (1:100, Alomone Labs) or against the extracellular N-terminal domain of rat mGluR1 (1:100, Alomone Labs) at 37°C for 6 minutes.

Techniques: Diffusion-based Assay, Control, Microscopy, Immunolabeling, Immunofluorescence

Journal: bioRxiv

Article Title: Single-molecule analysis reveals the rapid effect of estradiol on the surface movement of AMPAR in live neurons

doi: 10.1101/602813

Figure Lengend Snippet: Effect of estrogen receptor modulation on surface trafficking of AMPAR. ( a ) Representative PCR gel electrophoresis image depicting the expression of estrogen receptor mRNA (with a lack of expression of estrogen receptor alpha (ERα), estrogen receptor beta (ERβ), and G protein-coupled estrogen receptor 1 (GPER1)) in PC12 neurons. ( b ) Effect of E2 on the D value of mGluR1 (as the percent of vehicle treatment expressed as the median ± 25-75%, n= 840-860 trajectories per group). ( c ) Table showing the effect of ERβ agonist (DPN), GPER1 agonist (G1) and GPER1 antagonist (G15) treatment on D (µm 2 /s) values of GluR2-AMPAR molecules (expressed as the median ± 25-75% (in brackets), n=411-525 trajectories). ( d ) Table of the distributions of each minimal distance interaction type, including indirect (50-100 nm), microdomain (100-500 nm) and remote (500< nm), between GluR2-AMPAR and GPER1 on PC12 neurons after vehicle or E2 treatment (the minimal distance is expressed as the median ± 25-75% (in brackets)).

Article Snippet: Before single-molecule imaging, PC12 neurons were incubated in dRPMI with ATTO 488-labeled antibodies directed against the extracellular N-terminal domain of rat GluR2 (1:100, Alomone Labs) or against the extracellular N-terminal domain of rat mGluR1 (1:100, Alomone Labs) at 37°C for 6 minutes.

Techniques: Gel Electrophoresis, Expressing, Polymerase Chain Reaction

Journal: bioRxiv

Article Title: Single-molecule analysis reveals the rapid effect of estradiol on the surface movement of AMPAR in live neurons

doi: 10.1101/602813

Figure Lengend Snippet: The role of the cortical actin network in the rapid effect of E2. ( a ) The actin polymerization blocker latrunculin A (latA) decreased the dynamics of the cortical actin network in CHO cells (as % of control: 51 ± 11) (for the methods of analysis of cortical actin network dynamics, see Supplementary Methods). Neither 100 nM nor 100 pM E2 affected actin dynamics (as % of control: 100 pM E2 = 108 ± 30; 100 nM E2 = 93 ± 14) (n=5-6 cells per group) ( b ) Confocal images depicting the Alexa Fluor 488 phalloidin-labeled cortical actin network in PC12 neurons after vehicle or 10-minute 1 µM latA treatment in PC12 neurons. The bar graph shows the decreased integrated density of the cortical actin network after latA exposure (right, n=3 cells per group (3 regions of interest per cell)). Scale bar: 5 µm. ( c ). LatA treatment increased the diffusion coefficient (D) of GluR2-AMPA molecules on somas with no effect on the molecules on neurites (expressed as the median ± 25-75%, n = 307-507 trajectories). ( d ) On somas, 100 pM E2 decreased the D values of GluR2-AMPAR molecules. This effect was blocked in the presence of latA. Ineffectiveness of E2 action on neurite was not affected by latA treatment (expressed as the median ± 25-75%, n = 119-205 trajectories). **p<0.01, ***p<0.001

Article Snippet: Before single-molecule imaging, PC12 neurons were incubated in dRPMI with ATTO 488-labeled antibodies directed against the extracellular N-terminal domain of rat GluR2 (1:100, Alomone Labs) or against the extracellular N-terminal domain of rat mGluR1 (1:100, Alomone Labs) at 37°C for 6 minutes.

Techniques: Control, Analysis, Confocal, Labeling, Diffusion-based Assay

Journal: Nature

Article Title: Electrical and synaptic integration of glioma into neural circuits

doi: 10.1038/s41586-019-1563-y

Figure Lengend Snippet: a , Proliferative response of GFP (control) and GluA2-dominant negative subunit expressing glioma cells (GluA2-DN) after 24-hour exposure to soluble extracellular neuroligin-3 (NLGN3; 100nm) in the presence and absence of AMPA-receptor blocker, NBQX (10μM). b , Western blot analysis of phospho-AKT (Ser473) and total AKT in GFP (control) glioma cells in response to 5-minute exposure of soluble extracellular neuroligin-3 (NLGN3; 100nm) in the presence and absence of AMPA-receptor blocker, NBQX (10μM); left. Quantitative analysis of the ratio of pAKT/AKT normalized to vehicle (right). c , Western blot analysis of phospho-AKT (Ser473) and total AKT in GluA2-DN expressing glioma cells in response to 5-minute exposure of soluble extracellular neuroligin-3 (NLGN3; 100nm; left). Quantitative analysis of the ratio of pAKT/AKT normalized to vehicle (right). d , Time course of evoked glioma cell EPSC block by NASPM (100 μM, duration=red bar (n=7/5 cells/mice; left); Representative trace before (black) and after (red) addition of NASPM (right). e, Quantification of (d). f , GluA2 subunit Q/R editing efficiency in SU-DIPGXIII-FL and SU-DIPGVI cells as measured by PCR and expressed as % edited. g , Expression of ADAR1, the enzyme responsible for Q/R editing of GluA2 mRNA. Plot illustrates ADAR1 enzyme mRNA expression relative to beta-actin as measured by qPCR. Analyses in a,b,c,f,g were calculated from three independent sets of cells. Data shown as mean ± s.e.m. P values determined by one-way ANOVA with Tukey’s post-hoc analysis (a,b), by two-tailed Student’s t-test (c), by two-tailed paired Student’s t-test (e). All data shown as mean ± s.e.m. *P<0.01, **P<0.001, ***P<0.001, ****P<0.0001, NS = not significant.

Article Snippet: We introduced SpeI (5 ’ -TAAGCAactagtATGCAAAAGATTATGCAT-3’) and XmaI (5 ’ -TGCTTAcccgggCTAAATTTTAACACTTTCGAT-3’) restriction sites in full length human GRIA2 clone (GluA2 WT) obtained from Origene (RC212599) by PCR.

Techniques: Control, Dominant Negative Mutation, Expressing, Western Blot, Blocking Assay, Two Tailed Test

Journal: Nature

Article Title: Electrical and synaptic integration of glioma into neural circuits

doi: 10.1038/s41586-019-1563-y

Figure Lengend Snippet: a, Electrophysiological responses by model. Number of whole cell patch clamp recordings from cells in xenografted hippocampal slices separated by electrophysiological response to local electrical stimulation. b , Demonstration of depolarizing inward current in SU-DIPXIII-FL-ChR2 cells in response to single stimulation and 20Hz pulses of blue light as measured in current clamp (top) and voltage clamp (bottom). c , Proliferation index of xenografted SU-DIPGXIII-FL-YFP control glioma cells (no opsin expressed) in response to blue light stimulation or mock stimulation as measured by the proportion of GFP+/HNA+ cells expressing Ki67 24-hours after five optogenetic stimulation sessions (n=3 mice, mock stim; n=4 mice, stim). d, Quantification of cleaved caspase-3 in xenografted SU-DIPGXIII-FL-YFP control glioma cells in response to blue light stimulation or mock stimulation as measured by total number of HNA+ cells co-labeled with cleaved caspase-3 (n=3 mice/group). e , As in (d), quantification of cleaved caspase-3 in xenografted SU-DIPXIII-FL-ChR2 glioma cells (n=3 mice, mock stim; n=4 mice, stim). f , Validation of GluA2-dominative negative AMPA receptor subunit expressing construct. Representative traces of whole-cell voltage-clamp recording of WT (black) and GluA2-DN expressing (grey) SU-DIPGVI cells in response to 500μM (S)-AMPA (n=6 cells). g , Representative traces of whole-cell voltage-clamp recording in WT (black) and GluA2-DN expressing (grey) SU-DIPGXIII-FL cells in response to 500μM (S)-AMPA (n=6 cells). SU-DIPGXIII-FL cells are unable to homogeneously express the dominant construct, and therefore may be connected to WT GluA2 expressing cells, which accounts for the remaining current in the illustrated trace. Incorporation of the GluA2-DN construct thus results in a significantly abrogated AMPAR-dependent depolarization. Data shown as mean ± s.e.m for (c,d,e). All P-values determined by two-tailed Student’s t-test. NS = not significant.

Article Snippet: We introduced SpeI (5 ’ -TAAGCAactagtATGCAAAAGATTATGCAT-3’) and XmaI (5 ’ -TGCTTAcccgggCTAAATTTTAACACTTTCGAT-3’) restriction sites in full length human GRIA2 clone (GluA2 WT) obtained from Origene (RC212599) by PCR.

Techniques: Patch Clamp, Control, Expressing, Labeling, Biomarker Discovery, Construct, Two Tailed Test

Journal: Nature

Article Title: Electrical and synaptic integration of glioma into neural circuits

doi: 10.1038/s41586-019-1563-y

Figure Lengend Snippet: a , Optogenetic paradigm for glioma depolarization. ChR2-expressing glioma (blue), region of analysis (light blue). b , Proliferation index of SU-DIPGXIII-FL-ChR2 xenograft after mock stimulation (mock stim) or blue light stimulation (stim) measured as percent of GFP+/HNA+ cells expressing Ki67 (mock stim, n=8; stim, n=9 mice). c , As in (b), but SU-DIPGVI-ChR2 xenografts (n=6 mice/group). d , Representative confocal micrographs from (c), illustrating proliferating SU-DIPGVI-ChR2. Red=human nuclei; white=Ki67. Scale bar=50μm. e-f , Kaplan-Meier survival curves of SU-DIPGXIII-P* xenografts overexpressing e, GFP-only (green) or GluA2-WT-GFP (red) and f, GFP-only (in 80% of cells, green) or GluA2-DN-GFP (in 80% of cells, blue); n=5 mice/group. g , Competitive outgrowth of non-GluA2-DN-GFP-expressing cells in (f), determined by GFP/total human nuclei pixel intensity; (n=3 mice/group). h , Representative confocal micrographs of (f-g). White=human nuclei; green=GFP. Scale bar=50μm. i , Representative confocal images of SU-DIPGXIII-FL xenografts expressing GFP-only control (top) or GluA2-DN-GFP (bottom). Gray=MBP; White=glioma-GFP. Scale bar=500μm. j , Quantification of (i) (n=8 mice/group). k , Proliferation index of SU-DIPGVI xenografts treated with perampanel (AMPAR blocker) or vehicle control; (n=8 mice/group). l , Proliferation index of SU-DIPGXIII-FL in mice treated with meclofenamate (gap junction blocker) or vehicle control; (n=9 vehicle, n=8 treated mice). Data shown as mean±s.e.m (b,c,g,j,k,l). **P<0.01. ***P<0.001, ****P<0.0001. P-values determined by two-tailed unpaired Student’s t-test (b,c,g,k,l); two-tailed log rank analyses (e,f); two-sided Mann-Whitney test (j).

Article Snippet: We introduced SpeI (5 ’ -TAAGCAactagtATGCAAAAGATTATGCAT-3’) and XmaI (5 ’ -TGCTTAcccgggCTAAATTTTAACACTTTCGAT-3’) restriction sites in full length human GRIA2 clone (GluA2 WT) obtained from Origene (RC212599) by PCR.

Techniques: Expressing, Control, Two Tailed Test, MANN-WHITNEY

Journal: Nature

Article Title: Electrical and synaptic integration of glioma into neural circuits

doi: 10.1038/s41586-019-1563-y

Figure Lengend Snippet: a , Kaplan-Meier survival curves of second cohort of mice orthotopically xenografted with control GFP-only or GluA2-DN-GFP over-expressing cells (SU-DIPGXIII-P* xenograft model; n=5 mice per group). b , Representative coronal sections of mouse brains bearing SU-DIPGXIII-FL xenografts either expressing control GFP construct (left) or GluA2-DN-GFP construct; right). Gray, MBP; White, glioma-GFP. c , Proliferation indices of SU-DIPGXIII-FL cells at baseline in neuronal medium, in response to 10μM NBQX, in co-culture with neurons, or in co-culture with neurons in the presence of 10μM NBQX (n=3 biological replicates/group, except n=4 for baseline). d, Representative images of neuron-glioma co-cultures in the presence and absence of NBQX. Green = neurofilament (neuronal processes); Red = nestin (glioma cell processes); White = Ki67. Scale bar = 50μm. e , in vitro growth analysis of control GFP or GluA2-DN-GFP cells monitored over 3 days. f, in vitro apoptosis analysis of control GFP or GluA2-DN-GFP as measured by % of total cells co-stained with cleaved-caspase. g , 3D Matrigel invasion assay in WT (GFP) and GluA2-DN (GluA2-DN-GFP) expressing SU-DIPGXIII-FL cells 72 hours after seeding. h , Representative images of (g) at time 0 hr (left) and 72 hr (right) in control GFP-expressing (top) and GluA2-DN-GFP expressing cells (bottom). Scale bar = 1000μm. i , 3D migration assay in WT (GFP) and GluA2-DN (GluA2-DN-GFP) expressing SU-DIPGXIII-FL cells 72 hours after seeding. j , Representative images of (i) at time 0 hr (left) and 72 hr (right) in control GFP-expressing (top) and GluA2-DN-GFP expressing cells (bottom). Scale bar =1000μm. k, Representative confocal micrographs illustrating proliferating SU-DIPGVI cells in vehicle or perampanel-treated mice (n=8 mice/group). Red = human nuclei; white = Ki67. Scale bar = 50μm. l , IVIS bioluminescence analysis of overall tumor growth in SU-DIPGXIII-FL xenografts treated with vehicle or meclofenamate over a two-week period. Data represented as fold change in total flux; n=5 mice/group. Data shown as mean ± s.e.m. for (c,e,f,g,i,l). For analyses in (d-j), n=3 biological replicates. P-values determined by two-tailed log rank analyses (a), by one-way ANOVA with post-hoc analysis (c), by two-tailed unpaired Student’s t-test (f,g,i,l). *P<0.05, **P< 0.01,****P<0.0001. NS = not significant.

Article Snippet: We introduced SpeI (5 ’ -TAAGCAactagtATGCAAAAGATTATGCAT-3’) and XmaI (5 ’ -TGCTTAcccgggCTAAATTTTAACACTTTCGAT-3’) restriction sites in full length human GRIA2 clone (GluA2 WT) obtained from Origene (RC212599) by PCR.

Techniques: Control, Expressing, Construct, Co-Culture Assay, In Vitro, Staining, Invasion Assay, Migration, Two Tailed Test

Journal: Journal of Neurochemistry

Article Title: Targeting Glutamate Excitotoxicity With Memantine Modulates Glial Response and Protects Motoneurons After Spinal Root Lesion

doi: 10.1111/jnc.70429

Figure Lengend Snippet: Relative expression of genes involved in glutamatergic signaling in the different experimental groups ( n = 4/5 mice per group). The graphs demonstrate the analysis of the relative expression of the genes (A) Glast , (B) Glt1 , (C) Glul , (D) Gria2 . There was no significant difference in the expression of these genes after injury or treatment (one‐way ANOVA followed by Tukey's post‐test; mean ± SEM).

Article Snippet: Gria2 (GluA2) , Mm00442822_m1.

Techniques: Expressing