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rabbit anti-sult4a1 (Proteintech)

Name: rabbit anti-sult4a1
Brand: Proteintech
Rating: 90 (1 reviews)

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Proteintech rabbit anti-sult4a1
Rabbit Anti Sult4a1, supplied by Proteintech, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 90 stars, based on 1 article reviews

rabbit anti-sult4a1 - by Bioz Stars, 2026-03

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Physiologic expression of SULT4A1 protein during neuronal development. A, Representative Western blots of total cell lysates derived from rat cortical neurons at different days in vitro (n = 4 independent cultures for all conditions; one-way ANOVA test, p < 0.0001; Tukey's post hoc test, **p = 0.0011, ***p = 0.0001, ****p < 0.0001). B, Biochemical analysis of total brain lysates derived from wild-type male mice at different postnatal days (n = 3 animals for all conditions). C, Representative Western blots of protein lysates from H, S, Cx, and Cb derived from adult (P60) wild-type mice (n = 4 animals for all conditions; Kruskal–Wallis test, p < 0.0001; Dunn's post hoc test, *p = 0.0451; **p = 0.0065). D, Area-specific expression of SULT4A1 was also compared between adult M and F mice (n = 3 animals for all conditions; two-way ANOVA, p = 0.8362; Sidak's post hoc test). E, Representative immunocytochemical staining for SULT4A1 (green), βIII-Tubulin (red), and DAPI (blue), in rat cortical neurons at DIV1, DIV7, and DIV14. Scale bar, 50 μm. F, Left, Representative immunocytochemical staining of Oct4 (green), Sox2 (red), and DAPI (blue) of iPSCs obtained from healthy control individuals. Middle, iPSC-derived NSCs stained for Nestin (green), Sox2 (red), and DAPI (blue). Right, Representative immunostaining for MAP2 (green) and DAPI (blue) of NSC-derived neurons after 40 d of differentiation. Scale bar, 50 μm. G, Representative immunoblots of total cell lysates obtained from human neurons, sampled during differentiation from day 0 (NSC stage) until day 40 (mature neuron stage; n = 3 independent cultures for all conditions; one-way ANOVA, p < 0.0001; Tukey's post hoc test: 0 vs 30, p < 0.0001; 0 vs 40, p < 0.0001; 10 vs 20, p = 0.0371; 10 vs 30, p < 0.0001; 10 vs 40, p < 0.0001; 20 vs 30, p = 0.0003; 20 vs 40, p < 0.0001; 30 vs 40, p = 0.0411). Data represent the mean ± SEM.

Journal: The Journal of Neuroscience

Article Title: SULT4A1 Modulates Synaptic Development and Function by Promoting the Formation of PSD-95/NMDAR Complex

doi: 10.1523/JNEUROSCI.2194-19.2020

Figure Lengend Snippet: Physiologic expression of SULT4A1 protein during neuronal development. A, Representative Western blots of total cell lysates derived from rat cortical neurons at different days in vitro (n = 4 independent cultures for all conditions; one-way ANOVA test, p < 0.0001; Tukey's post hoc test, **p = 0.0011, ***p = 0.0001, ****p < 0.0001). B, Biochemical analysis of total brain lysates derived from wild-type male mice at different postnatal days (n = 3 animals for all conditions). C, Representative Western blots of protein lysates from H, S, Cx, and Cb derived from adult (P60) wild-type mice (n = 4 animals for all conditions; Kruskal–Wallis test, p < 0.0001; Dunn's post hoc test, *p = 0.0451; **p = 0.0065). D, Area-specific expression of SULT4A1 was also compared between adult M and F mice (n = 3 animals for all conditions; two-way ANOVA, p = 0.8362; Sidak's post hoc test). E, Representative immunocytochemical staining for SULT4A1 (green), βIII-Tubulin (red), and DAPI (blue), in rat cortical neurons at DIV1, DIV7, and DIV14. Scale bar, 50 μm. F, Left, Representative immunocytochemical staining of Oct4 (green), Sox2 (red), and DAPI (blue) of iPSCs obtained from healthy control individuals. Middle, iPSC-derived NSCs stained for Nestin (green), Sox2 (red), and DAPI (blue). Right, Representative immunostaining for MAP2 (green) and DAPI (blue) of NSC-derived neurons after 40 d of differentiation. Scale bar, 50 μm. G, Representative immunoblots of total cell lysates obtained from human neurons, sampled during differentiation from day 0 (NSC stage) until day 40 (mature neuron stage; n = 3 independent cultures for all conditions; one-way ANOVA, p < 0.0001; Tukey's post hoc test: 0 vs 30, p < 0.0001; 0 vs 40, p < 0.0001; 10 vs 20, p = 0.0371; 10 vs 30, p < 0.0001; 10 vs 40, p < 0.0001; 20 vs 30, p = 0.0003; 20 vs 40, p < 0.0001; 30 vs 40, p = 0.0411). Data represent the mean ± SEM.

Article Snippet: The following primary antibodies were used: mouse anti-β-actin (catalog #A5316, Sigma-Aldrich), mouse anti-βIII-tubulin (catalog #MAB1637, Sigma-Aldrich), mouse anti-GABA A Rα1 (catalog #75–136, NeuroMab), mouse anti-GAD65 (catalog #198111, Synaptic System), mouse anti-GAD67 (catalog #sc-28376, Santa Cruz Biotechnology), mouse anti-GFP (catalog #11814460001, Roche), rabbit anti-GluA1 (catalog #AB1504, Millipore), mouse anti-GluA2 (catalog #75–002, NeuroMab), mouse anti-GluN1 (catalog #556308, BD), mouse anti-GluN2A (catalog #75–288, NeuroMab), rabbit anti-GluN2B (catalog #06–600, Millipore), mouse anti-MAP2 (catalog #ab11268, Abcam), rabbit anti-mGlu5 (catalog #AB5675, Millipore), mouse anti-Nestin (catalog #MAB5326, Millipore), mouse anti-NLGN1 (catalog #75–160, NeuroMab), mouse anti-Oct4 (catalog #sc-5279, Santa Cruz Biotechnology), mouse anti-Pin1 (catalog #sc-46660, Santa Cruz Biotechnology), mouse anti-PSD-95 (catalog #75–028, NeuroMab), rabbit anti-PSD-95 (catalog #D27E11, Cell Signaling Technology), rabbit anti-Sox2 (catalog #11064–1-AP, Proteintech), rabbit anti-SULT4A1 (catalog #12578–1-AP, Proteintech), and rabbit anti-VGAT (catalog #131003, Synaptic System).

Techniques: Expressing, Western Blot, Derivative Assay, In Vitro, Staining, Immunostaining

Sult4a1 silencing reduces neuronal branching and dendritic spine density. A, Representative immunofluorescence images and relative traces of rat cortical neurons transfected with the scrambled shRNA (shCtrl), the Sult4a1-specific shRNA (shSult4a1), or shSult4a1 together with the resistant construct (Sult4a1r). Neurons were stained for SULT4A1 (red) to assess protein expression. Scale bar, 100 μm. B, Results of Sholl analysis and quantification of the number of intersections, plotted against the distance from the soma (shCtrl, n = 13 individual neurons; shSult4a1, n = 15 individual neurons; Sult4a1r, n = 19 individual neurons; two-way ANOVA, p < 0.0001; Tukey's post hoc test, *shSult4a1 vs shCtrl; #shSult4a1 vs Sult4a1r; 30 µm: ****p < 0.0001, ## p < 0.0001; 40 µm: ****p < 0.0001, ####p < 0.0001; 50 µm: ****p < 0.0001, ###p = 0.0001; 60 µm: ***p = 0.0007). C, Quantification of total dendrites length (shCtrl, n = 10 individual neurons; shSult4a1, n = 13 individual neurons; Sult4a1r, n = 13 individual neurons; one-way ANOVA, p < 0.0001; Tukey's post hoc test, shCtrl vs shSult4a1, p = 0.0001; shSult4a1 vs Sult4a1r, p = 0.0001; ns, not significant). D–E, Quantification of the number of primary (D) and secondary (E) dendrites [shCtrl, n = 21 individual neurons; shSult4a1, n = 18 individual neurons; Sult4a1r, n = 20 individual neurons; D: one-way ANOVA, p < 0.0001; Tukey's post hoc test: shCtrl vs shSult4a1, p = 0.0002; shSult4a1 vs Sult4a1r, p = 0.0002; ns, not significant; E: one-way ANOVA, p < 0.0001; Tukey's post hoc test, shCtrl vs shSult4a1, p < 0.0001; shSult4a1 vs Sult4a1r, p = 0.0079; ns, not significant]. F, Representative immunofluorescence images showing dendritic spines of rat cortical neurons transfected with shCtrl, shSult4a1, or shSult4a1 together with the resistant plasmid. Neurons were stained for SULT4A1 (red) to assess protein expression. Scale bar, 10 μm. G–I, Quantification of dendritic spines length (G), width (H), and number of spines per 10 μm of dendrite (I; shCtrl, n = 10 individual dendrites; shSult4a1, n = 10 individual dendrites; Sult4a1r, n = 14 individual dendrites; Kruskal–Wallis test: p = 0.6092 for G, p = 0.8472 for H, p = 0.0081 for I; I: Dunn's post hoc test: shCtrl vs shSult4a1, p = 0.0105; shSult4a1 vs Sult4a1r, p = 0.0417; ns, not significant). J, Quantification of filopodia, long thin and mushroom spines distribution. K, Representative immunohistochemistry images from brain slices obtained from 30-d-old in utero electroporated mice, showing cortical neurons expressing shCtrl or shSult4a1 and their relative traces. Scale bar, 100 μm. L, Results of Sholl analysis and quantification of the number of intersections, plotted against the distance from the soma (shCtrl, n = 10 individual neurons; shSult4a1, n = 13 individual neurons; two-way ANOVA, p = 0.0629; Sidak's post hoc test, 60 µm: *p = 0.0162; 70 µm: ****p < 0.0001). M, Quantification of total dendrites length (shCtrl, n = 10 individual neurons; shSult4a1, n = 13 individual neurons; unpaired t test, ***p = 0.0002). N, Representative immunofluorescence images showing dendritic spines of shCtrl- or shSult4a1-expressing cortical neurons from 30-d-old in utero electroporated mice. Scale bar, 10 μm. O–Q, Quantification of dendritic spines length (O), width (P), and number of spines per 10 μm of dendrite (Q; shCtrl, n = 12 individual dendrites; shSult4a1, n = 9 individual dendrites; unpaired t test p = 0.0490). R, Quantification of thin, stubby, and mushroom spines distribution. Data represent the mean ± SEM. shSult4a1 vs Sult4a1r, **p = 0.0079.

Journal: The Journal of Neuroscience

Article Title: SULT4A1 Modulates Synaptic Development and Function by Promoting the Formation of PSD-95/NMDAR Complex

doi: 10.1523/JNEUROSCI.2194-19.2020

Figure Lengend Snippet: Sult4a1 silencing reduces neuronal branching and dendritic spine density. A, Representative immunofluorescence images and relative traces of rat cortical neurons transfected with the scrambled shRNA (shCtrl), the Sult4a1-specific shRNA (shSult4a1), or shSult4a1 together with the resistant construct (Sult4a1r). Neurons were stained for SULT4A1 (red) to assess protein expression. Scale bar, 100 μm. B, Results of Sholl analysis and quantification of the number of intersections, plotted against the distance from the soma (shCtrl, n = 13 individual neurons; shSult4a1, n = 15 individual neurons; Sult4a1r, n = 19 individual neurons; two-way ANOVA, p < 0.0001; Tukey's post hoc test, *shSult4a1 vs shCtrl; #shSult4a1 vs Sult4a1r; 30 µm: ****p < 0.0001, ## p < 0.0001; 40 µm: ****p < 0.0001, ####p < 0.0001; 50 µm: ****p < 0.0001, ###p = 0.0001; 60 µm: ***p = 0.0007). C, Quantification of total dendrites length (shCtrl, n = 10 individual neurons; shSult4a1, n = 13 individual neurons; Sult4a1r, n = 13 individual neurons; one-way ANOVA, p < 0.0001; Tukey's post hoc test, shCtrl vs shSult4a1, p = 0.0001; shSult4a1 vs Sult4a1r, p = 0.0001; ns, not significant). D–E, Quantification of the number of primary (D) and secondary (E) dendrites [shCtrl, n = 21 individual neurons; shSult4a1, n = 18 individual neurons; Sult4a1r, n = 20 individual neurons; D: one-way ANOVA, p < 0.0001; Tukey's post hoc test: shCtrl vs shSult4a1, p = 0.0002; shSult4a1 vs Sult4a1r, p = 0.0002; ns, not significant; E: one-way ANOVA, p < 0.0001; Tukey's post hoc test, shCtrl vs shSult4a1, p < 0.0001; shSult4a1 vs Sult4a1r, p = 0.0079; ns, not significant]. F, Representative immunofluorescence images showing dendritic spines of rat cortical neurons transfected with shCtrl, shSult4a1, or shSult4a1 together with the resistant plasmid. Neurons were stained for SULT4A1 (red) to assess protein expression. Scale bar, 10 μm. G–I, Quantification of dendritic spines length (G), width (H), and number of spines per 10 μm of dendrite (I; shCtrl, n = 10 individual dendrites; shSult4a1, n = 10 individual dendrites; Sult4a1r, n = 14 individual dendrites; Kruskal–Wallis test: p = 0.6092 for G, p = 0.8472 for H, p = 0.0081 for I; I: Dunn's post hoc test: shCtrl vs shSult4a1, p = 0.0105; shSult4a1 vs Sult4a1r, p = 0.0417; ns, not significant). J, Quantification of filopodia, long thin and mushroom spines distribution. K, Representative immunohistochemistry images from brain slices obtained from 30-d-old in utero electroporated mice, showing cortical neurons expressing shCtrl or shSult4a1 and their relative traces. Scale bar, 100 μm. L, Results of Sholl analysis and quantification of the number of intersections, plotted against the distance from the soma (shCtrl, n = 10 individual neurons; shSult4a1, n = 13 individual neurons; two-way ANOVA, p = 0.0629; Sidak's post hoc test, 60 µm: *p = 0.0162; 70 µm: ****p < 0.0001). M, Quantification of total dendrites length (shCtrl, n = 10 individual neurons; shSult4a1, n = 13 individual neurons; unpaired t test, ***p = 0.0002). N, Representative immunofluorescence images showing dendritic spines of shCtrl- or shSult4a1-expressing cortical neurons from 30-d-old in utero electroporated mice. Scale bar, 10 μm. O–Q, Quantification of dendritic spines length (O), width (P), and number of spines per 10 μm of dendrite (Q; shCtrl, n = 12 individual dendrites; shSult4a1, n = 9 individual dendrites; unpaired t test p = 0.0490). R, Quantification of thin, stubby, and mushroom spines distribution. Data represent the mean ± SEM. shSult4a1 vs Sult4a1r, **p = 0.0079.

Techniques: Immunofluorescence, Transfection, shRNA, Construct, Staining, Expressing, Plasmid Preparation, Immunohistochemistry, In Utero

Sult4a1 overexpression (OV) impairs dendritic arborization and spines. A, Representative immunofluorescence images and relative traces of rat cortical neurons transfected with pLVTHM alone (GFP) or together with pFlag-SULT4A1 (overexpression). Neurons were stained for SULT4A1 (red) to assess protein (over)expression. Scale bar, 100 μm. B, Results of Sholl analysis and quantification of the number of intersections, plotted against the distance from the soma (GFP, n = 21 individual neurons; overexpression, n = 15 individual neurons; two-way ANOVA, p < 0.0001; Sidak's post hoc test: 10 µm, *p = 0.0126; 20 µm, *p = 0.0444). C, D, Quantification of the number of primary (C) and secondary (D) dendrites (GFP, n = 20 individual neurons; overexpression, n = 21 individual neurons; for C: Mann–Whitney test, p = 0.3511; for D: unpaired t test, p = 0.2496). E, Quantification of total dendrites length (GFP n = 20 individual neurons, Overexpression, n = 21 individual neurons; unpaired t test, p = 0.2387). F, Representative immunofluorescence images showing dendritic spines of rat cortical neurons transfected with pLVTHM alone (GFP) or together with pFlag-SULT4A1 (OV). Neurons were stained for SULT4A1 (red) to assess protein (over)expression. Scale bar, 10 μm. G–I, Quantification of dendritic spines length (G), width (H), and number of spines per 10 μm of dendrite (I; GFP, n = 15 individual dendrites; OV, n = 23 individual dendrites; unpaired t test, G, **p = 0.0126; I, *p = 0.0303). J, Quantification of filopodia, long thin and mushroom spines distribution. Data represent the mean ± SEM.

Journal: The Journal of Neuroscience

Article Title: SULT4A1 Modulates Synaptic Development and Function by Promoting the Formation of PSD-95/NMDAR Complex

doi: 10.1523/JNEUROSCI.2194-19.2020

Figure Lengend Snippet: Sult4a1 overexpression (OV) impairs dendritic arborization and spines. A, Representative immunofluorescence images and relative traces of rat cortical neurons transfected with pLVTHM alone (GFP) or together with pFlag-SULT4A1 (overexpression). Neurons were stained for SULT4A1 (red) to assess protein (over)expression. Scale bar, 100 μm. B, Results of Sholl analysis and quantification of the number of intersections, plotted against the distance from the soma (GFP, n = 21 individual neurons; overexpression, n = 15 individual neurons; two-way ANOVA, p < 0.0001; Sidak's post hoc test: 10 µm, *p = 0.0126; 20 µm, *p = 0.0444). C, D, Quantification of the number of primary (C) and secondary (D) dendrites (GFP, n = 20 individual neurons; overexpression, n = 21 individual neurons; for C: Mann–Whitney test, p = 0.3511; for D: unpaired t test, p = 0.2496). E, Quantification of total dendrites length (GFP n = 20 individual neurons, Overexpression, n = 21 individual neurons; unpaired t test, p = 0.2387). F, Representative immunofluorescence images showing dendritic spines of rat cortical neurons transfected with pLVTHM alone (GFP) or together with pFlag-SULT4A1 (OV). Neurons were stained for SULT4A1 (red) to assess protein (over)expression. Scale bar, 10 μm. G–I, Quantification of dendritic spines length (G), width (H), and number of spines per 10 μm of dendrite (I; GFP, n = 15 individual dendrites; OV, n = 23 individual dendrites; unpaired t test, G, **p = 0.0126; I, *p = 0.0303). J, Quantification of filopodia, long thin and mushroom spines distribution. Data represent the mean ± SEM.

Techniques: Over Expression, Immunofluorescence, Transfection, Staining, MANN-WHITNEY

Sult4a1 knockdown alters synaptic transmission. A, Representative immunoblots of total protein lysates derived from DIV14 rat cortical neurons transduced with a lentivirus expressing shCtrl or shSult4a1. Both conditions were compared with not infected condition (Ni) so as to verify whether the infection itself could cause any alteration of Sult4a1 protein expression. Sult4a1 protein levels were normalized against the level of the not infected neurons (Ni, n = 7 independent cultures; shCtrl, n = 4 independent cultures; shSult4a1, n = 7 independent cultures; one-sample t test: Ni vs shSult4a1, ****p < 0.0001; shCtrl vs shSult4a1, ****p < 0.0001; ns, not significant). B, Representative Western blots of total lysates from shCtrl- or shSult4a1-transduced cortical neurons. Protein levels were normalized against the level of the shCtrl-transduced neurons (shCtrl, n ≥ 3 independent cultures; shSult4a1, n ≥ 3 independent cultures; one-sample t test: GAD65, *p = 0.0396; GluN1, *p = 0.0186). C, Top panels, Representative sEPSCs recorded from shCtrl-transfected (blue) or shSult4a1-transfected (red) neurons. Middle panels, left, Plot of cumulative probability of sEPSC frequencies (shCtrl, n = 5 individual neurons; shSult4a1, n = 5 individual neurons; Kolmogorov–Smirnov test, p = 4 × 10−18); right, mean instantaneous frequencies plot (shCtrl, n = 5 individual neurons; shSult4a1, n = 5 individual neurons; Mann–Whitney test, *p < 0.05; data are shown as the mean ± SEM). Bottom panels, left, Plot of cumulative probability of sEPSC amplitudes (Kolmogorov–Smirnov test, p = 10−11); right, plot of mean sEPSC amplitudes (shCtrl, n = 5 individual neurons; shSult4a1, n = 5 individual neurons; Mann–Whitney test, p = 10−12; data are shown as the mean ± SEM). D, Top panels, Representative sIPSCs recorded from shCtrl-transfected (blue) or shSult4a1-transfected (red) neurons. Middle panels, Left, Plot of cumulative probability of sIPSC frequencies (shCtrl, n = 5 individual neurons; shSult4a1, n = 6 individual neurons; Kolmogorov–Smirnov test, p = 5 × 10−8); right, plot of mean instantaneous frequencies (shCtrl, n = 5 individual neurons; shSult4a1, n = 6 individual neurons; Mann–Whitney test; data are shown as the mean ± SEM). Bottom panels, Left, Plot of cumulative probability of sIPSC amplitudes (Kolmogorov–Smirnov test, p = 10−12); right, plot of mean sIPSC amplitudes (shCtrl, n = 5 individual neurons; shSult4a1, n = 5 individual neurons; Mann–Whitney test, p = 10−8; data are shown as the mean ± SEM).

Journal: The Journal of Neuroscience

Article Title: SULT4A1 Modulates Synaptic Development and Function by Promoting the Formation of PSD-95/NMDAR Complex

doi: 10.1523/JNEUROSCI.2194-19.2020

Figure Lengend Snippet: Sult4a1 knockdown alters synaptic transmission. A, Representative immunoblots of total protein lysates derived from DIV14 rat cortical neurons transduced with a lentivirus expressing shCtrl or shSult4a1. Both conditions were compared with not infected condition (Ni) so as to verify whether the infection itself could cause any alteration of Sult4a1 protein expression. Sult4a1 protein levels were normalized against the level of the not infected neurons (Ni, n = 7 independent cultures; shCtrl, n = 4 independent cultures; shSult4a1, n = 7 independent cultures; one-sample t test: Ni vs shSult4a1, ****p < 0.0001; shCtrl vs shSult4a1, ****p < 0.0001; ns, not significant). B, Representative Western blots of total lysates from shCtrl- or shSult4a1-transduced cortical neurons. Protein levels were normalized against the level of the shCtrl-transduced neurons (shCtrl, n ≥ 3 independent cultures; shSult4a1, n ≥ 3 independent cultures; one-sample t test: GAD65, *p = 0.0396; GluN1, *p = 0.0186). C, Top panels, Representative sEPSCs recorded from shCtrl-transfected (blue) or shSult4a1-transfected (red) neurons. Middle panels, left, Plot of cumulative probability of sEPSC frequencies (shCtrl, n = 5 individual neurons; shSult4a1, n = 5 individual neurons; Kolmogorov–Smirnov test, p = 4 × 10−18); right, mean instantaneous frequencies plot (shCtrl, n = 5 individual neurons; shSult4a1, n = 5 individual neurons; Mann–Whitney test, *p < 0.05; data are shown as the mean ± SEM). Bottom panels, left, Plot of cumulative probability of sEPSC amplitudes (Kolmogorov–Smirnov test, p = 10−11); right, plot of mean sEPSC amplitudes (shCtrl, n = 5 individual neurons; shSult4a1, n = 5 individual neurons; Mann–Whitney test, p = 10−12; data are shown as the mean ± SEM). D, Top panels, Representative sIPSCs recorded from shCtrl-transfected (blue) or shSult4a1-transfected (red) neurons. Middle panels, Left, Plot of cumulative probability of sIPSC frequencies (shCtrl, n = 5 individual neurons; shSult4a1, n = 6 individual neurons; Kolmogorov–Smirnov test, p = 5 × 10−8); right, plot of mean instantaneous frequencies (shCtrl, n = 5 individual neurons; shSult4a1, n = 6 individual neurons; Mann–Whitney test; data are shown as the mean ± SEM). Bottom panels, Left, Plot of cumulative probability of sIPSC amplitudes (Kolmogorov–Smirnov test, p = 10−12); right, plot of mean sIPSC amplitudes (shCtrl, n = 5 individual neurons; shSult4a1, n = 5 individual neurons; Mann–Whitney test, p = 10−8; data are shown as the mean ± SEM).

Techniques: Transmission Assay, Western Blot, Derivative Assay, Transduction, Expressing, Infection, Transfection, MANN-WHITNEY

$SULT4A1 interaction with Pin1 and its role in excitatory synapses. A, Left, Representative INMDA traces elicited by the perfusion of 100 μm NMDA from neurons transfected with shCtrl (blue), shSult4a1 (red), or shSult4a1 together with the resistant plasmid Sult4a1r (orange). Right, Analysis of the mean peak INMDA current densities in neurons transfected with shCtrl, shSult4a1, or shSult4a1 together with the resistant plasmid Sult4a1r (shCtrl, n = 30 individual neurons; shSult4a1, n = 34 individual neurons; Sult4a1r, n = 10; one-way ANOVA, p = 0.0028; Tukey's post hoc test: shCtrl vs shSult4a1, p = 0.0048; shSult4a1 vs Sult4a1r, p = 0.0423; ns, not significant). shCtrl vs shSult4a1, **p = 0.0048; shSult4a1 vs Sult4a1r, *p = 0.0423. B, Left, Representative IAMPA traces elicited by the perfusion of 100 μm AMPA from neurons transfected with shCtrl (blue), shSult4a1 (red), or shSult4a1 together with the resistant plasmid Sult4a1r (orange). Right, Analysis of mean peak IAMPA current densities in neurons transfected with shCtrl, shSult4a1, or shSult4a1 together with the resistant plasmid Sult4a1r (shCtrl, n = 11 individual neurons; shSult4a1, n = 8 individual neurons; Sult4a1r, n = 7 individual neurons; one-way ANOVA, p = 0.2133). C, Representative immunofluorescence images showing dendritic spines of rat cortical neurons transfected with shCtrl or shSult4a1. Neurons were stained for PSD-95 (blue) and GluN1 (red). Scale bar, 10 μm. D, E, Quantification of the number of PSD-95 (D) and GluN1 (E) clusters per 10 μm of dendrite (shCtrl, n = 11 individual dendrites; shSult4a1, n = 11 individual dendrites; unpaired t test, E, p = 0.0003). F, Quantification of GluN1 and PSD-95 clusters colocalization (shCtrl, n = 11 individual dendrites; shSult4a1, n = 11, individual dendrites; unpaired t test, ***p = 0.0004). G, Representative Western blots of synaptosomal fractions obtained from cortical neurons transduced with shCtrl or shSult4a1. Protein levels were normalized against the levels of the shCtrl-transduced neurons (shCtrl, n ≥ 3 independent cultures; shSult4a1, n ≥ 3 independent cultures; one-sample t test, SULT4A1, p < 0.0001; GluN1, p = 0.0032; Pin1, p = 0.0206). H, Left, Representative images of immunoprecipitation (IP) assay performed on synaptosomal fractions derived from neurons transduced with shCtrl or shSult4a1. Proteins were precipitated using mouse anti-Pin1 or mouse IgG antibodies, and nitrocellulose membranes were probed with anti-Pin1, anti-SULT4A1, and anti-PSD-95 antibodies (WB). Right, Histogram showing the ratio between PSD-95 and Pin1 signals (PSD-95/Pin1; shCtrl, n = 4 independent experiments; shSult4a1, n = 4 independent experiments; unpaired t test, p = 0.0464). Data represent the mean ± SEM. *p = 0.0464.$

Journal: The Journal of Neuroscience

Article Title: SULT4A1 Modulates Synaptic Development and Function by Promoting the Formation of PSD-95/NMDAR Complex

doi: 10.1523/JNEUROSCI.2194-19.2020

Figure Lengend Snippet: SULT4A1 interaction with Pin1 and its role in excitatory synapses. A, Left, Representative INMDA traces elicited by the perfusion of 100 μm NMDA from neurons transfected with shCtrl (blue), shSult4a1 (red), or shSult4a1 together with the resistant plasmid Sult4a1r (orange). Right, Analysis of the mean peak INMDA current densities in neurons transfected with shCtrl, shSult4a1, or shSult4a1 together with the resistant plasmid Sult4a1r (shCtrl, n = 30 individual neurons; shSult4a1, n = 34 individual neurons; Sult4a1r, n = 10; one-way ANOVA, p = 0.0028; Tukey's post hoc test: shCtrl vs shSult4a1, p = 0.0048; shSult4a1 vs Sult4a1r, p = 0.0423; ns, not significant). shCtrl vs shSult4a1, **p = 0.0048; shSult4a1 vs Sult4a1r, *p = 0.0423. B, Left, Representative IAMPA traces elicited by the perfusion of 100 μm AMPA from neurons transfected with shCtrl (blue), shSult4a1 (red), or shSult4a1 together with the resistant plasmid Sult4a1r (orange). Right, Analysis of mean peak IAMPA current densities in neurons transfected with shCtrl, shSult4a1, or shSult4a1 together with the resistant plasmid Sult4a1r (shCtrl, n = 11 individual neurons; shSult4a1, n = 8 individual neurons; Sult4a1r, n = 7 individual neurons; one-way ANOVA, p = 0.2133). C, Representative immunofluorescence images showing dendritic spines of rat cortical neurons transfected with shCtrl or shSult4a1. Neurons were stained for PSD-95 (blue) and GluN1 (red). Scale bar, 10 μm. D, E, Quantification of the number of PSD-95 (D) and GluN1 (E) clusters per 10 μm of dendrite (shCtrl, n = 11 individual dendrites; shSult4a1, n = 11 individual dendrites; unpaired t test, E, p = 0.0003). F, Quantification of GluN1 and PSD-95 clusters colocalization (shCtrl, n = 11 individual dendrites; shSult4a1, n = 11, individual dendrites; unpaired t test, ***p = 0.0004). G, Representative Western blots of synaptosomal fractions obtained from cortical neurons transduced with shCtrl or shSult4a1. Protein levels were normalized against the levels of the shCtrl-transduced neurons (shCtrl, n ≥ 3 independent cultures; shSult4a1, n ≥ 3 independent cultures; one-sample t test, SULT4A1, p < 0.0001; GluN1, p = 0.0032; Pin1, p = 0.0206). H, Left, Representative images of immunoprecipitation (IP) assay performed on synaptosomal fractions derived from neurons transduced with shCtrl or shSult4a1. Proteins were precipitated using mouse anti-Pin1 or mouse IgG antibodies, and nitrocellulose membranes were probed with anti-Pin1, anti-SULT4A1, and anti-PSD-95 antibodies (WB). Right, Histogram showing the ratio between PSD-95 and Pin1 signals (PSD-95/Pin1; shCtrl, n = 4 independent experiments; shSult4a1, n = 4 independent experiments; unpaired t test, p = 0.0464). Data represent the mean ± SEM. *p = 0.0464.

Techniques: Transfection, Plasmid Preparation, Immunofluorescence, Staining, Western Blot, Transduction, Immunoprecipitation, Derivative Assay

Rescue of shSult4a1-dependent phenotypes via Pin1 pharmacological inhibition. A, Left, Representative INMDA traces from shCtrl-transfected (blue) and shSult4a1-transfected (red) neurons, treated for 48 h with 2.5 μm PiB or vehicle. Right, Analysis of mean peak INMDA current densities (shCtrl + vehicle, n = 12 individual neurons; shCtrl + PiB, n = 16 individual neurons; shSult4a1 + vehicle, n = 6 individual neurons; shSult4a1 + PiB, n = 16 individual neurons; one-way ANOVA, p = 0.0082; Tukey's post hoc test: shCtrl vehicle vs shSult4a1 vehicle, p = 0.0146; shSult4a1 vehicle vs shSult4a1 PiB, p = 0.0186). shCtrl vehicle vs shSult4a1 vehicle, *p = 0.0146; shSult4a1 vehicle vs shSult4a1 PiB, *p = 0.0186. B, Left, Representative INMDA traces from shCtrl-transfected (blue) and shSult4a1-transfected (red) neurons, treated for 48 h with 4 μm DTM or vehicle. Right, Analysis of mean peak INMDA current densities (shCtrl + vehicle, n = 16 individual neurons; shCtrl + DTM, n = 7 individual neurons; shSult4a1 + vehicle, n = 21 individual neurons; shSult4a1 + DTM, n = 10 individual neurons; one-way ANOVA, p = 0.0008; Tukey's post hoc test: shCtrl + vehicle vs shSult4a1 + vehicle, p = 0.0455; shSult4a1 + vehicle vs shSult4a1 + DTM, p = 0.0006). shCtrl vehicle vs shSult4a1 vehicle, *p = 0.0455; shSult4a1 vehicle vsshSult4a 1 DTM, ***p = 0.0006. C, Left, Representative immunofluorescence images showing dendritic spines of rat cortical neurons transfected with shCtrl or shSult4a1 and treated for 48 h with 2.5 μm PiB, a pharmacological inhibitor of Pin1 catalytic activity, or vehicle (DMSO). Scale bar, 10 μm. Right, Quantification of the number of dendritic spines per 10 μm of dendrite (shCtrl + vehicle, n = 19 individual dendrites; shCtrl + PiB, n = 16 individual dendrites; shSult4a1 + vehicle, n = 19 individual dendrites; shSult4a1 + PiB, n = 17 individual dendrites; one-way ANOVA, p = 0.0003; Tukey's post hoc test: shCtrl + vehicle vs shSult4a1 + vehicle, p = 0.0096; shCtrl + PiB vs shSult4a1 vehicle, p = 0.0005; shSult4a1 + vehicle vs shSult4a1 + PiB, p = 0.0044). shCtrl vehicle vs shSult4a1 vehicle, **p = 0.0096; shCtrl PiB vs shSult4a vehicle, ***p = 0.0005; shSult4avehicle vs shSult4a1 PiB **p = 0.0044. D, Left, Representative immunofluorescence images showing dendritic spines of rat cortical neurons transfected with shCtrl or shSult4a1 and treated at DIV12 for 48 h with 4 μm DTM, a pharmacological inhibitor of Pin1 catalytic activity, or vehicle (DMSO). Scale bar, 10 µm. Right, Quantification of the number of dendritic spines per 10 μm of dendrite (shCtrl + vehicle, n = 10 individual dendrites; shCtrl + DTM, n = 10 individual dendrites; shSult4a1 + vehicle, n = 11 individual dendrites; shSult4a1 + DTM, n = 10 individual dendrites; Kruskal–Wallis test, p = 0.0006; Dunn's post hoc test: shCtrl + vehicle vs shSult4a1 + vehicle, p = 0.0301; shCtrl + DTM vs shSult4a1 vehicle, p = 0.0012; shSult4a1 + vehicle vs shSult4a1 + DTM, p = 0.0005). shCtrl vehicle vs shSult4a1 vehicle, *p = 0.0301; shCtrl DTM vs shSult4a1 vehicle, **p = 0.0012; shSult4a1 vehicle vs shSult4a1 DTM,**p = 0.0005). E, Cartoon showing the hypothesized mechanism. SULT4A1 interacts with Pin1 promoting the formation of the PSD-95/NMDAR complex in excitatory synapses. In the absence of SULT4A1, Pin1 is free to interact with PSD-95, reducing NMDAR expression in synapses. Data represent the mean ± SEM.

Journal: The Journal of Neuroscience

Article Title: SULT4A1 Modulates Synaptic Development and Function by Promoting the Formation of PSD-95/NMDAR Complex

doi: 10.1523/JNEUROSCI.2194-19.2020

Figure Lengend Snippet: Rescue of shSult4a1-dependent phenotypes via Pin1 pharmacological inhibition. A, Left, Representative INMDA traces from shCtrl-transfected (blue) and shSult4a1-transfected (red) neurons, treated for 48 h with 2.5 μm PiB or vehicle. Right, Analysis of mean peak INMDA current densities (shCtrl + vehicle, n = 12 individual neurons; shCtrl + PiB, n = 16 individual neurons; shSult4a1 + vehicle, n = 6 individual neurons; shSult4a1 + PiB, n = 16 individual neurons; one-way ANOVA, p = 0.0082; Tukey's post hoc test: shCtrl vehicle vs shSult4a1 vehicle, p = 0.0146; shSult4a1 vehicle vs shSult4a1 PiB, p = 0.0186). shCtrl vehicle vs shSult4a1 vehicle, *p = 0.0146; shSult4a1 vehicle vs shSult4a1 PiB, *p = 0.0186. B, Left, Representative INMDA traces from shCtrl-transfected (blue) and shSult4a1-transfected (red) neurons, treated for 48 h with 4 μm DTM or vehicle. Right, Analysis of mean peak INMDA current densities (shCtrl + vehicle, n = 16 individual neurons; shCtrl + DTM, n = 7 individual neurons; shSult4a1 + vehicle, n = 21 individual neurons; shSult4a1 + DTM, n = 10 individual neurons; one-way ANOVA, p = 0.0008; Tukey's post hoc test: shCtrl + vehicle vs shSult4a1 + vehicle, p = 0.0455; shSult4a1 + vehicle vs shSult4a1 + DTM, p = 0.0006). shCtrl vehicle vs shSult4a1 vehicle, *p = 0.0455; shSult4a1 vehicle vsshSult4a 1 DTM, ***p = 0.0006. C, Left, Representative immunofluorescence images showing dendritic spines of rat cortical neurons transfected with shCtrl or shSult4a1 and treated for 48 h with 2.5 μm PiB, a pharmacological inhibitor of Pin1 catalytic activity, or vehicle (DMSO). Scale bar, 10 μm. Right, Quantification of the number of dendritic spines per 10 μm of dendrite (shCtrl + vehicle, n = 19 individual dendrites; shCtrl + PiB, n = 16 individual dendrites; shSult4a1 + vehicle, n = 19 individual dendrites; shSult4a1 + PiB, n = 17 individual dendrites; one-way ANOVA, p = 0.0003; Tukey's post hoc test: shCtrl + vehicle vs shSult4a1 + vehicle, p = 0.0096; shCtrl + PiB vs shSult4a1 vehicle, p = 0.0005; shSult4a1 + vehicle vs shSult4a1 + PiB, p = 0.0044). shCtrl vehicle vs shSult4a1 vehicle, **p = 0.0096; shCtrl PiB vs shSult4a vehicle, ***p = 0.0005; shSult4avehicle vs shSult4a1 PiB **p = 0.0044. D, Left, Representative immunofluorescence images showing dendritic spines of rat cortical neurons transfected with shCtrl or shSult4a1 and treated at DIV12 for 48 h with 4 μm DTM, a pharmacological inhibitor of Pin1 catalytic activity, or vehicle (DMSO). Scale bar, 10 µm. Right, Quantification of the number of dendritic spines per 10 μm of dendrite (shCtrl + vehicle, n = 10 individual dendrites; shCtrl + DTM, n = 10 individual dendrites; shSult4a1 + vehicle, n = 11 individual dendrites; shSult4a1 + DTM, n = 10 individual dendrites; Kruskal–Wallis test, p = 0.0006; Dunn's post hoc test: shCtrl + vehicle vs shSult4a1 + vehicle, p = 0.0301; shCtrl + DTM vs shSult4a1 vehicle, p = 0.0012; shSult4a1 + vehicle vs shSult4a1 + DTM, p = 0.0005). shCtrl vehicle vs shSult4a1 vehicle, *p = 0.0301; shCtrl DTM vs shSult4a1 vehicle, **p = 0.0012; shSult4a1 vehicle vs shSult4a1 DTM,**p = 0.0005). E, Cartoon showing the hypothesized mechanism. SULT4A1 interacts with Pin1 promoting the formation of the PSD-95/NMDAR complex in excitatory synapses. In the absence of SULT4A1, Pin1 is free to interact with PSD-95, reducing NMDAR expression in synapses. Data represent the mean ± SEM.

Techniques: Inhibition, Transfection, Immunofluorescence, Activity Assay, Expressing

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