EMBO/soda-nel huggingface dataset:
An inter-dimer allosteric switch controls NMDA receptor activity (2018)
Figure 1

Figure 1: NMDAR activation requires conformational mobility at inter-domain and inter-layer interfaces.

(A) Schematic representation of a full-length GluN1/GluN2B NMDAR. The receptor displays a layered architecture. NTD, N-terminal domain; ABD; agonist-binding domain; TMD, transmembrane domain. The studied inter-domain and inter-layer interfaces are reported with numbered yellow stars. For clarity, interface 4 is shown from a side view (inset).

(B) Representative current traces from oocytes expressing wild-type (WT) and double cysteine mutant NMDARs before and after DTE treatment. For each mutant, traces are normalized (in height, not in current amplitude) to the maximal response obtained after DTE treatment on wild-type (WT) receptors.

(C) Summary of the DTE-induced potentiation of WT and double cysteine mutants. Mean and n values are given in Appendix Table S1. ***P<0.001, one-way ANOVA on ranks (Kruskal-Wallis H test) followed by Bonferroni corrected Dunn's test.

(D) Immunoblots from Xenopus oocytes expressing either wt or mutant subunits. M1 indicates the GluN1 monomer (~110 kDa); M2 the GluN2B monomer (~180 kDa); D1/1 the GluN1 homodimer (~220 kDa); and D1/2 the GluN1/GluN2B heterodimer (~290 kDa). Lower panels: immunoblots in reducing conditions (+ β-mercaptoethanol). N.I., non-injected oocytes.
An inter-dimer allosteric switch controls NMDA receptor activity (2018)
Figure 2

Figure 2: Rolling between the two constitutive ABD dimers boosts receptor activity.

(A) Left, localization of site 5 at the interface between the two constitutive ABD dimers. Right top, crystal structures of the inhibited (PDB 5IOV; Zhu et al, 2016) and presumably active (5FXG; Tajima et al, 2016) states illustrating the differences in distances between GluN2B-L795 and GluN1-E698 or GluN1-R673 at interface 5. Those three residues are colored green, while GluN1 and GluN2B subunits are colored red and blue respectively. Right bottom, schematic representation of the GluN1-R673C/GluN2B-L795C disulfide cross-link capturing an inhibited state of the receptor, and of the GluN1-E698C/GluN2B-L795C disulfide cross-link trapping the rolling motion.

(B) Assessment of receptor channel activity using MK-801 inhibition kinetics. Representative current traces from oocytes expressing either wild-type (WT) or GluN1-E698C/GluN2B-L795C mutant receptors in response to 10 nM MK-801 during agonist application. Responses were scaled to the current amplitude obtained before MK-801 application. Inset, mono-exponential fits of MK-801 wash-in and wash-out. Note the strikingly faster kinetics in mutant receptors, both at the onset and offset of MK-801.

(C) Relative MK-801 inhibition on- and off-rate constants (kon and koff). All values were normalized to the value obtained for WT GluN1/GluN2B receptors. Mean and n values are given in Appendix Table S2. ***P<0.001, one-way ANOVA on ranks followed by Bonferroni corrected Dunn's test. Error bars, SD.
An inter-dimer allosteric switch controls NMDA receptor activity (2018)
Figure 3

Figure 3: Super-active receptors are insensitive to NTD-mediated allosteric modulation.

(A) Schematic representation of the super-active GluN1-E698C/GluN2B-L795C receptor locked in the 'rolled' conformation, showing the NTD binding sites for spermine, zinc and ifenprodil, positive (+) and negative (-) allosteric modulators, respectively. Open circles indicate that the binding site is on the other side of the GluN2B NTD. The allosteric inhibitor H+ is also indicated, although its binding site remains ill-defined.

(B) Zinc, ifenprodil and pH dose-response curves of wild-type (WT) GluN1/GluN2B receptors and mutant GluN1-E698C/GluN2B-L795C receptors. For comparison, zinc and ifenprodil dose-response curves of GluN1/GluN2B receptors lacking the whole GluN2B NTD (GluN1 WT/GluN2B-delNTD) are also shown (dashed lines; data from Rachline et al, 2005). Values of IC50, maximal inhibition, Hill coefficient and n are given in Appendix Table S3. Error bars, SD.

(C) Spermine (200 µM, pH 6.3) potentiation of WT GluN1/GluN2B and mutant GluN1-E698C/GluN-L795C receptors. Spermine (200 µM, pH 6.5) sensitivity is also shown for WT GluN1/GluN2B receptors and receptors lacking either the GluN1 (GluN1-delNTD/GluN2B WT) or GluN2B (GluN1 WT/GluN2B-delNTD) NTD (data from Mony et al, 2011). Mean values and n are given in Appendix Table S3. **P<0.01, one-way ANOVA on ranks followed by Bonferroni corrected Dunn's test. Error bars, SD.

(D) Glutamate and glycine dose-response curves of WT GluN1/GluN2B receptors and mutant GluN1-E698C/GluN2B-L795C receptors. Values of EC50, Hill coefficient and n are given in Appendix Table S3. Error bars, SD.
An inter-dimer allosteric switch controls NMDA receptor activity (2018)
Figure 5

Figure 5: Structural mobility of an inter-layer GluN1 protruding loop during rolling.

(A) Left, localization of interface 1 between GluN1 ABD and GluN2B NTD. This interface involves a GluN1 specific loop that protrudes from GluN1 ABD upper lobe towards GluN2B NTD lower lobe. Right, Initial and final frames of the fitting presented in Fig 4 at interface 1. The two pairs of residues targeted for cysteine mutations GluN1-N492/GluN2B-S188 and GluN1-K495/GluN2B-N192 are shown as spheres linked by a dotted line colored black and red, respectively.

(B) Evolution of Cα-Cα distances of the two pairs of residues GluN1-N492/GluN2B-S188 and GluN1-K495/GluN2B-N192 during the fitting.

(C) MK-801 inhibition kinetics. On-rate constants (kon) of inhibition by 10 nM MK-801 on wild-type (WT) and mutant receptors. All values are normalized to that obtained for WT GluN1/GluN2B receptors. Values of Mean and n are given in Appendix Table S2. *P<0.05, one-way ANOVA on ranks followed by Bonferroni corrected Dunn's test. Error bars, SD.
An inter-dimer allosteric switch controls NMDA receptor activity (2018)
Figure 6

Figure 6: Influence of rolling on single-receptor gating kinetics.

(A) Representative recordings of patches containing one single wild-type (WT) or GluN1-E698C/GluN2B-L795C receptor. The bottom trace is an expanded view. C, closed channel; O, open channel.

(B) Single channel properties (equilibrium channel open probability, mean open time, mean closed time) of the WT and super-active GluN1-E698C/GluN2B-L795C receptors. Mean and n values are given in Appendix Table S4. *P<0.05, two-tailed Student's t-test, unpaired. Error bars, SEM.

(C) Closed (left) and open (right) time histograms for WT (upper panels) or super-active GluN1-E698C/GluN2B-L795C (lower panels) receptors. Closed time histograms were best fit with 5 exponentials, whereas open time histograms were best fit with 2 exponentials (see Online Methods and Appendix Fig S8). Smooth lines are associated exponential fits.

(D) Kinetic schemes and equilibrium constants for WT (upper panel) or super-active GluN1-E698C/GluN2B-L795C (lower panels) receptors. Records were analyzed at equilibrium. Ligand-binding steps are shown in gray. Note that for simplicity, long-lived desensitized steps are not shown. *P<0.05 relative to WT GluN1/GluN2B (two-tailed Student's t-test, unpaired).

(E) Proposed mechanism for the conformational switch in full-length tetrameric GluN1/GluN2B receptor: inter-dimer rolling in the ABD layer is coupled to entry of the NTD layer in its active state. The positive allosteric modulator spermine, which binds the interface between GluN1 and GluN2B NTD lower lobes, enhances receptor activity by stabilizing the NTD compact form and therefore rolling. Conversely, the negative allosteric modulators zinc and ifenprodil inhibit receptor activity by stabilizing an expanded form of the NTDs (lower lobes further apart) thus preventing rolling. The rolling motion facilitates pore opening by acting on the ABD-TMD connecting linkers.
Gut Microbiota Influences Pathological Angiogenesis in Obesity-driven Choroidal Neovascularization (2016)
Figure 2

Figure 2. High-fat diet increases recruitment of microglia and macrophages.

(A-C) 3D rendering of Z-stack confocal image showing Isolectin B4-stained laser-burn with FITC-dextran-labeled CNV and IBA-1-stained MPs. (D) Representative FACS plots of retinas and sclera-choroid-RPE-cell complexes from Regular-chow Diet (RD)-fed mice without and 3, 7 and 14 days after laser-burn. (E) Quantification of MPs (Ly6G -, F4/80+, CD11b+) at p3, p7 and p14; fold naïve (no burn); n= 5(no burn), 5(p3), 6(p7), 5(p14); CI 95%. (F) Representative FACS plots and (G) quantification of microglia (Ly6G-, F4/80+, CD11b+, CX3CR1hi CD45lo) fold naïve; n= 5(no burn), 4(p3), 6(p7), 5(p14); CI 95%. (H) Representative FACS plots of retinas and sclera-choroid-RPE-cell complexes from RD- and HFD-fed mice, with vehicle or antibiotic treatment at p7. (I) Quantification of MPs at p7 fold RD; n= 6(RD), 4(HFD), 3(HFD+AB), 4(RD+AB); CI 95%. (J) Representative FACS plots and (K) quantification of microglia (Ly6G-, F4/80+, CD11b+, CX3CR1hi CD45lo) fold RD; n= 5(RD), 5(HFD), 3(HFD+AB), 4(RD+AB); CI 95%. (L) Representative confocal images of IBA-1-stained MPs on choroidal flat mounts from RD- and HFD-fed mice, treated with vehicle or with neomycin. Examples of labeled macrophages (white dots) are presented in side-panels. (M) Total number of MPs around laser impact area; n= 7(RD), 9(HFD), 4(HFD+AB), 7(RD+AB), with 23(RD), 29(HFD), 13(HFD+AB), 23(RD+AB) burns total; CI 99%. All comparisons between groups are analyzed using one-way analysis of variance (ANOVA) and Tukey’s multiple comparisons test; *P< 0.05, **P< 0.01, ***P< 0.001; error bars represent mean ± S.E.M. Each “n” represents one mouse per experimental group; CI: Confidence interval. Scale bar: 100μm.
Gut Microbiota Influences Pathological Angiogenesis in Obesity-driven Choroidal Neovascularization (2016)
Figure 3

Figure 3. High-fat diet augments gut permeability, metabolic endotoxemia and systemic inflammation.

(A) Concentration of gut absorbed Evans Blue in serum 24 hours after oral administration in Regular-chow Diet (RD)- and high-fat diet (HFD)-fed mice, treated with vehicle or with antibiotic treatment; n= 6(RD), 5(HFD), 5(HFD+AB), 5(RD+AB); CI 99%. (B) Activation of PRRs induced by stimulation with serum isolated from mice fed RD or HFD for 7 weeks and receiving vehicle or neomycin for 3 weeks; fold RD; n=3 for all groups; CI 99%. (C) Serum cytokine profiles determined by Bio-Plex assay (Bio-Rad) of IL-1β, n= 11(RD), 12(HFD), 12(HFD+AB), 11(RD+AB); CI 95%; IL-6, n= 11(RD), 12(HFD), 12(HFD+AB), 11(RD+AB), CI 99.9%; TNF-α n=11 for all groups, CI 99%; IL-17A n= 11(RD), 12(HFD), 12(HFD+AB), 11(RD+AB); CI 95%; and IFN-γ; n= 12 for all groups; CI 95%. (D) mRNA expression of Il-1β; n= 5(RD), 7(HFD), 7(HFD+AB), 3(RD+AB); CI 95%, Il-6; n= 5(RD), 5(HFD), 4(HFD+AB), 4(RD+AB); CI 95%, Tnf-α; n= 6(RD), 9(HFD), 4(HFD+AB), 5(RD+AB); CI 95% and Vegf-a; n= 6(RD), 5(HFD), 5(HFD+AB), 6(RD+AB); CI 95% in choroids. All comparisons between groups are analyzed using one-way analysis of variance (ANOVA) and Tukey’s multiple comparisons test; *P< 0.05, **P< 0.01, ***P< 0.001; error bars represent mean ± S.E.M. Each “n” represents one mouse; CI: Confidence interval.
Gut Microbiota Influences Pathological Angiogenesis in Obesity-driven Choroidal Neovascularization (2016)
Figure 4

Figure 4. High-fat diet exacerbates CNV through gut microbiota.

(A) Schematic representation of microbiotal transfer experiments where recipient mice are gavaged with a suspension of fecal pellets in PBS from donor mice that are either fed a Regular-chow Diet (RD) or a high-fat diet (HFD). (B) Experimental timeline describing preparation of mice for microbiotal transfers where mice receive 5 days of antibiotics (Neomycin and Ampicillin) at 6 weeks of age. Starting at 7 weeks of life, RD x RDT- and HFD x RDT-mice receive weekly microbiotal transplants from RD donor mice and HFD x HFDT-mice receive weekly microbiotal transplants from HFD donor mice until sacrifice at week 13. At the age of 11 weeks, mice are subjected to 4 laser burns per eye. (C) Weight gain and (D) area under the curve of percentage weight gain of RD x RDT, HFD x RDT and HFD x HFDT mice; n= 13(RDxRDT), 12(HFDxHFDT), 14(HFDxRDT); CI 99%. (E) Representative circle charts of relative abundance of bacterial phyla in gut microbiota of RD-fed mice receiving a RD transfer and HFD-fed mice receiving a HFD or RD transfer (F) relative proportion per experimental group of different phyla; n= 5(RDxRDT), 5(HFDxHFDT), 4(HFDxRDT); Bacteroidetes; CI 99%, Firmicutes; CI 95%, Proteobacteria; CI 95%, Actinobacteria and Spirochaetes. (G) Compilation of compressed Z-stack confocal images of FITC-dextran-labeled CNV and single-plane confocal images of isolectin B4-stained choroidal flat mounts from RD x RDT, HFD x RDT and HFD x HFDT mice. (H) Quantification of area of FITC-dextran-labeled CNV over area of isolectin B4-stained laser impact area fold RD x RDT; n= 6(RDxRDT), 9(HFDxHFDT), 9(HFDxRDT), with 15(RDxRDT), 21(HFDxHFDT), 16(HFDxRDT) burns total; CI 95%. (I) Concentration of gut absorbed Evans Blue in serum 24 hours after oral administration in RD- and HFD-fed mice, after RD or HFD microbiotal transplant; n= 3(RDxRDT), 3(HFDxHFDT), 4(HFDxRDT); CI 95%. (J) mRNA expression of Il-1β; n= 5(RDxRDT), 4(HFDxHFDT), 5(HFDxRDT), Il-6; n= 5(RDxRDT), 4(HFDxHFDT), 4(HFDxRDT); CI 99%, Tnf-α; n= 5(RDxRDT), 4(HFDxHFDT), 4(HFDxRDT); CI 95% and Vegf-a; n= 5(RDxRDT), 4(HFDxHFDT), 5(HFDxRDT); CI 95% in choroids. All comparisons between groups are analyzed using one-way analysis of variance (ANOVA) and Tukey’s multiple comparisons test; *P< 0.05, **P< 0.01, ***P< 0.001; error bars represent mean ± S.E.M. Each “n” represents one mouse, per experimental group, in (I) both eyes were used for mRNA extraction, giving two data points per mouse; CI: Confidence interval. Scale bar: 100μm.
A highly potent and selective Vps34 inhibitor alters vesicle trafficking and autophagy (2014)
figf1

(a) Chemical structure of SAR405. (b) SAR405 inhibits the function of Vps34 in the GFP-FYVE cellular assay. HeLa cells stably transfected with GFP-FYVE were incubated with DMSO (left) or with 1 μM SAR405 (right) for 2 h. Cells were fixed, and nuclei were stained using Hoechst 33342 (blue). Fluorescence was analyzed using an imaging cytometer (X40). One representative image out of three independent experiments is depicted. Scale bars, 20 μm. (c) SAR405 does not prevent phosphorylation of Akt and S6. U87MG cells were treated with SAR405 or PI103 for 2 h at the indicated concentrations. The effect of compounds on phosphorylated Akt (p-Akt) Ser473 and Thr308 residues, total Akt and on phosphorylated S6 (Ser240 and Ser244 residues) and total S6 protein was measured using western blotting. One representative experiment from at least two independent experiments is shown. Full-length blots are shown in Supplementary Figure 6.
A highly potent and selective Vps34 inhibitor alters vesicle trafficking and autophagy (2014)
figf3

(a) SAR405 induces late endosome-lysosome swelling. RKO cells were treated with DMSO (upper panels) or with 10 μM SAR405 for 16 h (lower panels). The late endosomes-lysosomes compartment was assessed using Lysotracker green (left) or Alexa 488–conjugated anti-Lamp1 antibodies (right), as indicated. Nuclei were stained using Hoechst 33342 (blue). One representative image chosen from three independent experiments, showing fluorescence imaged using a Zeiss microscope (X40), is shown for each condition. Arrows indicate accumulation of swollen late endosome-lysosomes. Scale bars, 20 μm. (b) SAR405 triggers accumulation of p62. RKO cells were treated with the indicated concentration of Vps34 inhibitor for 24 h. The effect on the p62 biomarker was analyzed by western blotting. GAPDH was used as a loading control. The results are representative of at least two independent experiments. Full-length blots are shown in Supplementary Figure 14. (c) SAR405 affects the maturation of cathepsin D (cat D). RKO cells were treated at the indicated concentrations of SAR405 for 24 h. The different forms of cathepsin D (PPCD, pre-pro-catD) were analyzed by western blotting. GAPDH was used as a loading control. The results are representative of at least two independent experiments. Full-length blots are shown in Supplementary Figure 16.
A highly potent and selective Vps34 inhibitor alters vesicle trafficking and autophagy (2014)
figf4

(a) GFP-LC3 HeLa cells were cultured in fed conditions or starved using EBSS plus 10 μM hydroxychloroquine and treated with Vps34 inhibitor. Cells were fixed, and nuclei were stained using Hoechst 33342 (blue). One representative image, acquired with an iCyte imaging cytometer, for three independent experiments is depicted (X40). White arrows indicate GFP-LC3 spots, which correspond to the autophagosomes. Scale bars, 10 μm. (b) Histograms show the percentage of positive cells and GFP intensity, measured using an imaging cytometer for different compound concentrations on GFP-LC3 HeLa cells. Black and gray histograms correspond to fed and starved conditions, respectively. For GFP intensity, the values were normalized to 1 for fed conditions. Data show mean ± s.d. for three independent experiments. (c) Wild-type HeLa and H1299 cell lines were cultured either in fed or starved conditions with EBSS and 10 μM hydroxychloroquine (HCQ), as indicated. Cells were treated at the same time with SAR405 at different concentrations. LC3-II was measured by western blotting, and GAPDH was used as a loading control. One representative experiment for three independent experiments is shown. Full-length blots are shown in Supplementary Figure 18. (d) Histograms show the LC3-II/GAPDH ratio for each condition in the two cell lines. Black and gray histograms correspond to fed condition and EBSS conditions, respectively. The addition of hydroxychloroquine at 10 μM is indicated. Values were normalized to 1 for fed conditions. Data show mean ± s.d. for three independent experiments.
A highly potent and selective Vps34 inhibitor alters vesicle trafficking and autophagy (2014)
figf5

(a) GFP-LC3 H1299 cells were treated with 1 μM of the catalytic mTOR inhibitor AZD8055 in fed conditions with DMSO (left) or with 1 μM SAR405 (right). Cells were fixed, and nuclei were stained using Hoechst 33342 (blue). One representative image of cells out of three independent experiments, acquired using an iCyte imaging cytometer, is shown (X40). White arrows indicate the GFP-LC3 spots, which correspond to the autophagosomes. Scale bars, 10 μm. (b) Histograms show the percentage of positive cells (top) and the GFP intensity (bottom), as measured using an imaging cytometer. Treatment of GFP-LC3 H1299 in fed conditions with 1 μM of AZD8055 and different concentrations of SAR405 is indicated. For GFP intensity, the values were normalized to 1 for fed conditions. Data show mean ± s.d. for three independent experiments. (c) The concentration needed to reach 40% inhibition (IC40) of cell proliferation is indicated. The top histograms indicate the IC40 of SAR405 as a single agent and in combination with everolimus in the three cell lines. The bottom histograms indicate the IC40 of everolimus as a single agent and in combination with SAR405. The IC40 values for the combination of the two compounds were determined using rays with an effective fraction ∼0.5, corresponding to compounds that are in equipotent proportion in the mixture (Supplementary Fig. 20). The absolute IC40 determination of both compounds is represented as the geometric mean from two or three independent experiments.
Bipolar-associated miR-499-5p controls neuroplasticity by downregulating the Cav1.2 subunit CACNB2 (2022)
Figure 1

Figure 1: miR-499-5p is expressed in rat hippocampal pyramidal neurons and functions as a negative regulator of dendritogenesis.

miR-499-5p qPCR analysis using total RNA isolated from different areas of the adult female rat brain. U6 snRNA was used for normalization. Data are represented as bar graphs, mean±S.D., (n=3 animals).

miR-499-5p qPCR analysis using total RNA isolated from the hippocampus of male rats that were either group-housed or socially isolated for 4 weeks post-weaning. Data are represented as box plot with whiskers (+: mean, line: median; whiskers: Tukey). (n=9 rats per group; Mann-Whitney U-test; *** p=0.0008). Fold changes represent changes in gene expression relative to the control condition. U6 snRNA was used for normalization.

Representative picture of single-molecule fluorescence in situ hybridization (smFISH) performed in rat hippocampal neurons at DIV7 using probes directed against miR-499-5p (red channel) and CamK2a (green channel) to identify excitatory neurons. Scale bar: 20 mm.

Representative grey-scale images of primary rat hippocampal neurons (DIV 10) transfected with GFP and the indicated miRNA mimics. Scale bars: 50 μm.

Quantification of the mean number of intersections by Sholl analysis. GFP-only transfected conditions were set to one in each experiment. Data are represented as scattered dot plots with bar, mean±S.D. (n=3; Unpaired two-sample t-Test, *p=0.0435).

Representative grey-scale images of GFP-transfected Cacna1c+/+ (WT) or Cacna1c+/- primary rat hippocampal neurons (DIV 10). Scale bars = 50 μm.

Quantification of the mean number of intersections by Sholl analysis. Data are represented as scattered dot plots with bar, mean±S.D. (n=3 independent experiments; Unpaired two-sample t-Test, **p=0.0096). Fold changes represent changes in dendritic complexity relative to the control condition.
Bipolar-associated miR-499-5p controls neuroplasticity by downregulating the Cav1.2 subunit CACNB2 (2022)
Figure 2

Figure 2: The BD risk gene Cacbn2 is a downstream target of miR-499-5p in the regulation of hippocampal neuron dendritogenesis.

Nucleotide base pairing of miR-499-5p with Cacnb2 3'UTR (left) and schematic representation of the Cacnb2 luciferase reporter (right).

qPCR analysis of Cacnb2 mRNA levels using total RNA isolated from rat hippocampal neurons transfected with either a control or miR-499-5p mimic. Data are represented as scattered dot plots with bar, mean±S.D. (n=3 independent experiments; Unpaired two-sample t-test, *p=0.0363).

qPCR analysis of Cacnb2 mRNA levels using total RNA from the hippocampus of either socially isolated or group-housed rats. Data are represented as box plot with whiskers (+: mean, line: median; whiskers: Tukey). (n=9 rats per group; Unpaired two-sample t-test, **p=0.0020).

Relative luciferase activity of rat hippocampal neurons transfected with the indicated miRNA mimics and expressing either a Cacnb2 WT or MUT reporter. Data are represented as scattered dot plots with bar, mean±S.D. (n=4 independent experiments; Two-way ANOVA: main effect of the miRNA mimic p=0.0087, of the Cacnb2 luciferase reporter p<0.0001 and of the miRNA mimic by Cacnb2 luciferase reporter interaction p<0.0001, Tukey's HSD: ***p=0.0002).

Relative luciferase activity of rat hippocampal neurons transfected with the indicated pLNAs and expressing either a Cacnb2 WT or MUT reporter. Data are represented as scattered dot plots with bar, mean±S.D. (n=4 independent experiments; Two-way ANOVA: main effect of the miRNA pLNA p=0.0061, no main effect of the Cacnb2 luciferase reporter p=0.3590 or the miRNA pLNA by Cacnb2 luciferase reporter p=0.0910, Tukey's HSD: *p=0.0152). Fold changes represent changes in relative luciferase activity of transfected neurons relative to the non-transfected neurons.

Representative images of DIV 10 hippocampal neurons co-transfected with control or miR-499-5p mimics and a pMT2-CACNB2 expression plasmid (CACNB2). Scale bars = 50 μm.

Quantification of the mean number of intersections by Sholl analysis. Data are represented as scattered dot plots with bar, mean±S.D. (n=3 independent experiments; Two-way ANOVA: no main effect of the CACNB2 transfection p=0.5027 or the miRNA mimics p=0.0805, main effect of the CACBN2 transfection by miRNA mimics interaction p=0.0213, Tukey's HSD: *p=0.0303). Fold changes represent the changes in dendritic complexity of transfected neurons compared to non-treated control neurons. ns = not significant.