Journal: Neurochemistry international
Article Title: Sexually dimorphic and brain region-specific transporter adaptations in system x c - null mice.
doi: 10.1016/j.neuint.2020.104888
Figure Lengend Snippet: Fig. 1. Comparison of glutamate and cysteine transporter levels between wild-type (þ/þ) and system xc - null (sut/sut) female mice in striatum, hippo campus, and cortex. Plasma membrane striatal, hippocampal, and cortical protein extracted in pairs from +/+ and sut/sut female mice [5 striata or 5 hippocampi (5 mice per 2 samples), or 2 cortices (1 mouse per sample)] were separated under reducing conditions via 8% SDS-PAGE. Representative blots and protein quantification for EAAT1, 2, 3, ASCT1, SNAT3 and LAT2 protein in striatum (A), hippocampus (B) and cortex (C). Protein quantification was performed by normalizing to each samples’ representative total protein to correct for differences in loading followed by baseline correction to the paired +/+ levels. Values are graphed as Tukey’s box and whisker plots with the box representing the 2nd and 3rd quartiles, and whiskers representing points within 1.5 times the interquartile distance. An asterisk (*) represents a significant between-group difference calculated using a ratio-paired t-test on transformed data. Between-group differences for each brain areas are as follows: (A) Striatum: EAAT1 (p = 0.044; n = 6), EAAT2 (p = 0.477; n = 11), EAAT3 (p = 0.234; n = 10), ASCT1 (p = 0.343; n = 14), SNAT3 (p = 0.279; n = 6) and LAT2 (p = 0.376; n = 6). (B) Hippocampus: EAAT1 (p = 0.085; n = 11), EAAT2 (p = 0.169; n = 11), EAAT3 (p = 0.086; n = 7), ASCT1 (p = 0.144; n = 14), SNAT3 (p = 0.213; n = 7) and LAT2 (p = 0.436; n = 7). (C) Cortex: EAAT1 (p = 0.092; n = 14), EAAT2 (p = 0.2391; n = 14), EAAT3 (p = 0.426; n = 14), ASCT1 (p = 0.0267; n = 13), SNAT3 (p = 0.0001; n = 6), and LAT2 (p = 0.260; n = 6).
Article Snippet: Protein Host Species Company Catalog Number RRID Epitope EAAT1 Rabbit Abcam Ab416 AB_304334 C-terminus of rat EAAT1 EAAT2 Rabbit Abcam Ab41621 AB_941789 C-terminus of rat EAAT2 EAAT3 Rabbit Abcam Ab124802 AB_10974334 Human EAAT3 residues 150-250 ASCT1 Rabbit Alomone Labs ANT-081 AB_2756719 Mouse Slc1a4 residues 500-512 LAT2 Mouse OriGene TA500514 AB_11124222 Full length Slc7a8 from HEK293T cells SNAT3 Rabbit Proteintech 14315-1-AP AB_941782 Residues 1–191 of Slc38a3 fusion protein SOD1 Rabbit Millipore 07–043 AB_310587 C-terminus of human Cu/Zn SOD SOD2 Rabbit Sigma Aldrich HPA001814 AB_1080134 Residues 2–146 of human SOD2 Catalase Mouse Sigma Aldrich C0979 AB_258720 Human erythrocyte catalase H.M. Sosnoski et al. Neurochemistry International 141 (2020) 104888 comparisons that controlled for procedural variabilities in the amount of protein extracted.
Techniques: Comparison, Clinical Proteomics, Membrane, SDS Page, Whisker Assay, Transformation Assay
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![Fig. 1. Comparison of glutamate and cysteine transporter levels between wild-type (þ/þ) and system xc - null (sut/sut) female mice in striatum, hippo campus, and cortex. Plasma membrane striatal, hippocampal, and cortical protein extracted in pairs from +/+ and sut/sut female mice [5 striata or 5 hippocampi (5 mice per 2 samples), or 2 cortices (1 mouse per sample)] were separated under reducing conditions via 8% SDS-PAGE. Representative blots and protein quantification for EAAT1, 2, 3, <t>ASCT1,</t> SNAT3 and LAT2 protein in striatum (A), hippocampus (B) and cortex (C). Protein quantification was performed by normalizing to each samples’ representative total protein to correct for differences in loading followed by baseline correction to the paired +/+ levels. Values are graphed as Tukey’s box and whisker plots with the box representing the 2nd and 3rd quartiles, and whiskers representing points within 1.5 times the interquartile distance. An asterisk (*) represents a significant between-group difference calculated using a ratio-paired t-test on transformed data. Between-group differences for each brain areas are as follows: (A) Striatum: EAAT1 (p = 0.044; n = 6), EAAT2 (p = 0.477; n = 11), EAAT3 (p = 0.234; n = 10), <t>ASCT1</t> (p = 0.343; n = 14), SNAT3 (p = 0.279; n = 6) and LAT2 (p = 0.376; n = 6). (B) Hippocampus: EAAT1 (p = 0.085; n = 11), EAAT2 (p = 0.169; n = 11), EAAT3 (p = 0.086; n = 7), ASCT1 (p = 0.144; n = 14), SNAT3 (p = 0.213; n = 7) and LAT2 (p = 0.436; n = 7). (C) Cortex: EAAT1 (p = 0.092; n = 14), EAAT2 (p = 0.2391; n = 14), EAAT3 (p = 0.426; n = 14), ASCT1 (p = 0.0267; n = 13), SNAT3 (p = 0.0001; n = 6), and LAT2 (p = 0.260; n = 6).](https://pub-med-unpaywalled-images-cdn.bioz.com/pub_med_ids_ending_with_9267/pm33199267/pm33199267__page3_image1.jpg)
![Fig. 2. Comparison of glutamate and cysteine transporter levels between wild-type (þ/þ) and system xc - null (sut/sut) male mice in striatum, hippo campus, and cortex. Plasma membrane striatal, hippocampal, and cortical protein extracted in pairs from +/+ and sut/sut male mice [5 striata or 5 hippocampi (5 mice per 2 samples), or 2 cortices (1 mouse per sample)] were separated under reducing conditions via 8% SDS-PAGE. Representative blot for detection of EAAT1, 2, 3, ASCT1, SNAT3 and LAT2 protein in striatum (A), hippocampus (B), and cortex (C). Protein quantification was performed by normalizing to each samples’ representative total protein levels to correct for differences in loading followed by baseline correction to the paired +/+ levels. Values are graphed as Tukey’s box and whisker plots with the box representing the 2nd and 3rd quartiles, and whiskers representing points within 1.5 times the interquartile distance. An asterisk (*) represents a significant between-group difference calculated using a ratio-paired t-test on transformed data. (A) Striatum: EAAT1 (p = 0.3866; n = 7), EAAT2 (p = 0.468; n = 7), EAAT3 (p = 0.351; n = 6), ASCT1 (p = 0.493; n = 7), SNAT3 (p = 0.0267; n = 7) and LAT2 (p = 0.465; n = 7). (B) Hippocampus: EAAT1 (p = 0.186; n = 7), EAAT2 (p = 0.079; n = 7), EAAT3 (p = 0.487; n = 6), ASCT1 (p = 0.469; n = 7), SNAT3 (p = 0.438; n = 7) and LAT2 (p = 0.386; n = 6). (C) Cortex: EAAT1 (p = 0.149; n = 7), EAAT2 (p = 0.161; n = 7), EAAT3 (p = 0.325; n = 7), ASCT1 (p = 0.429; n = 7), SNAT3 (p = 0.026; n = 5) and LAT2 (p = 0.254; n = 5).](https://pub-med-unpaywalled-images-cdn.bioz.com/pub_med_ids_ending_with_9267/pm33199267/pm33199267__page4_image1.jpg)