anti girk2 (Alomone Labs)


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Anti Girk2, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/anti girk2/product/Alomone Labs
Average 93 stars, based on 1 article reviews
Price from $9.99 to $1999.99
Images
1) Product Images from "One-shot design elevates functional expression levels of a voltage-gated potassium channel"
Article Title: One-shot design elevates functional expression levels of a voltage-gated potassium channel
Journal: bioRxiv
doi: 10.1101/2022.12.28.522065

Figure Legend Snippet: (a) Currents of the paddle chimera (PC) and designs are plotted against varying test voltages. Each curve represents recordings from n ⩾8 oocytes. Error bars represent standard deviations. (b) Average currents (paddle chimera: 0.91±0.19 μA, D1: 4.65±1.52 μA, D2: 12.50±1.63 μA). (c) Western blot results reporting total membrane expression. The blots were labeled with anti-FLAG (top) or with anti-GIRK2 (as control; bottom) antibodies. (d) Normalized blot intensities. The experiment was performed in triplicate. Error bars represent the standard deviation. (e) Normalized tail currents were recorded at -50mV as a function of the test pulse voltage (see inset). Measured tail current amplitudes were fitted with a single-component Boltzmann equation (solid lines), from which V 1/2 of activation was inferred. (f) Inferred V 1/2 values (paddle chimera: 16.10±5.39 mV, D1: -8.33±3.49 mV, and, D2: -16.61±2.75 mV).
Techniques Used: Western Blot, Expressing, Labeling, Standard Deviation, Activation Assay
anti girk2 (Alomone Labs)


Structured Review

Anti Girk2, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/anti girk2/product/Alomone Labs
Average 93 stars, based on 1 article reviews
Price from $9.99 to $1999.99
Images
1) Product Images from "One-shot design elevates functional expression levels of a voltage-gated potassium channel"
Article Title: One-shot design elevates functional expression levels of a voltage-gated potassium channel
Journal: bioRxiv
doi: 10.1101/2022.12.28.522065

Figure Legend Snippet: (a) Currents of the paddle chimera (PC) and designs are plotted against varying test voltages. Each curve represents recordings from n ⩾8 oocytes. Error bars represent standard deviations. (b) Average currents (paddle chimera: 0.91±0.19 μA, D1: 4.65±1.52 μA, D2: 12.50±1.63 μA). (c) Western blot results reporting total membrane expression. The blots were labeled with anti-FLAG (top) or with anti-GIRK2 (as control; bottom) antibodies. (d) Normalized blot intensities. The experiment was performed in triplicate. Error bars represent the standard deviation. (e) Normalized tail currents were recorded at -50mV as a function of the test pulse voltage (see inset). Measured tail current amplitudes were fitted with a single-component Boltzmann equation (solid lines), from which V 1/2 of activation was inferred. (f) Inferred V 1/2 values (paddle chimera: 16.10±5.39 mV, D1: -8.33±3.49 mV, and, D2: -16.61±2.75 mV).
Techniques Used: Western Blot, Expressing, Labeling, Standard Deviation, Activation Assay
rabbit anti girk2 (Alomone Labs)


Structured Review

Rabbit Anti Girk2, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/rabbit anti girk2/product/Alomone Labs
Average 93 stars, based on 1 article reviews
Price from $9.99 to $1999.99
Images
1) Product Images from "Temporal-spatial changes in Sonic Hedgehog expression and signaling reveal different potentials of ventral mesencephalic progenitors to populate distinct ventral midbrain nuclei"
Article Title: Temporal-spatial changes in Sonic Hedgehog expression and signaling reveal different potentials of ventral mesencephalic progenitors to populate distinct ventral midbrain nuclei
Journal: Neural Development
doi: 10.1186/1749-8104-6-29

Figure Legend Snippet: Cells labeled with Shh-GIFM at different time points have a changing potential to contribute to subpopulations of DA neurons . (A-H) Immunofluorescent staining for β-gal-positive fate-mapped cells (Shh-GIFM at E8.5 or E11.5) and DA neurons (TH) on coronal sections of the ventral midbrain (P21 to P30). The areas shown in (A-D) are indicated in (E-H). Arrows indicate double-labeled cells; arrowheads indicate β-gal-positive cells with astrocytic morphology. (E'-H') Representative schematics of the immunostained sections showing the distribution of TH-positive fate-mapped cells (red dots) and of fate-mapped cells with astrocytic morphology (yellow crosses). Rostral, Bregma -2.92; caudal, Bregma -3.40 . If, interfascicular nucleus; Pn, paranigral nucleus; Snc, substantia nigra pars compacta; Snl, substantia nigra lateralis; Vta, VTA. Fate-mapped cells outside these areas are not represented. Cells with astrocytic morphology are not present with TM8.5. Scale bars: (A-D) 40 μm; (E-H) 200 μm. (I) Relative contribution of cells marked with Shh-GIFM between E8.5 and E11.5 to the SN (Snl + Snc), dorsal-lateral VTA (Vta) and ventral-medial VTA (Pn + If); see schematic in (L). For each animal (n ≥ 3), TH-positive fate-mapped cells were counted in the three indicated areas and normalized for the combined number of overlapping cells counted in these areas (in percent). Error bars indicate standard deviation. Significance (* P < 0.05; ** P < 0.01; *** P < 0.001) was determined by ANOVA and LSD post-hoc analysis. (J) Distribution of Calbindin- and Girk2-positive cells. (K) Relative contribution of fate-mapped cells to Calbindin (VTA) versus Girk2 (SN) positive cells. Calbindin- or Girk2-positive fate-mapped cells were counted in three different rostral-caudal midbrain areas (n ≥ 3). The ratio of Calbindin-positive fate-mapped cells to Girk2-positive fate-mapped cells was determined. Significance (*** P < 0.001) was determined by Student's t -test. (L) Schematic showing the SN, dlVTA and vmVTA. (M) Fate mapping strategy.
Techniques Used: Labeling, Staining, Standard Deviation
girk2 (Alomone Labs)


Structured Review

Girk2, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/girk2/product/Alomone Labs
Average 86 stars, based on 1 article reviews
Price from $9.99 to $1999.99
Images
1) Product Images from "Spatial Memory Training Counteracts Hippocampal GIRK Channel Decrease in the Transgenic APP Sw,Ind J9 Alzheimer’s Disease Mouse Model"
Article Title: Spatial Memory Training Counteracts Hippocampal GIRK Channel Decrease in the Transgenic APP Sw,Ind J9 Alzheimer’s Disease Mouse Model
Journal: International Journal of Molecular Sciences
doi: 10.3390/ijms232113444

Figure Legend Snippet: Effect of genotype and age on hippocampal levels of GIRK channel subunits and modulator. Protein levels of GIRK channel subunits (1–4) and modulator (RGS7) in the hippocampus of non-transgenic (WT) and APP Sw,Ind J9 mice at 6 and 12–18 months of age. ( A ) GIRK1, ( B ) GIRK2, ( C ) GIRK3, ( D ) GIRK4 and ( E ) RGS7. Data is expressed as percentage relative to 6-month-old WT mice (100%). ** p < 0.01, *** p < 0.001 vs. WT 6-month-old; # p < 0.05 vs. APP Sw,Ind J9 6-month-old. m, months; βAct, β actin.
Techniques Used: Transgenic Assay

Figure Legend Snippet: Effect of genotype and spatial memory training on hippocampal levels of GIRK channel subunits and modulator. Protein levels of GIRK channel subunits (1–4) and modulator (RGS7) in the hippocampus of naïve and memory-trained non transgenic (WT) and APP Sw,Ind J9 mice at 6 months of age. ( A ) GIRK1, ( B ) GIRK2, ( C ) GIRK3, ( D ) GIRK4 and ( E ) RGS7. Data is expressed as percentage relative to untrained naïve WT mice (100%). ** p < 0.01 vs. naïve WT. βAct, β actin.
Techniques Used: Transgenic Assay

Figure Legend Snippet: Characteristics of primary and secondary antibodies used to measure protein expression by Western Blot.
Techniques Used: Expressing, Western Blot
apc 006 (Alomone Labs)


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Apc 006, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/apc 006/product/Alomone Labs
Average 93 stars, based on 1 article reviews
Price from $9.99 to $1999.99
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rabbit anti girk2 (Alomone Labs)


Structured Review

Rabbit Anti Girk2, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/rabbit anti girk2/product/Alomone Labs
Average 93 stars, based on 1 article reviews
Price from $9.99 to $1999.99
Images
1) Product Images from "Alcohol reverses the effects of KCNJ6 (GIRK2) noncoding variants on excitability of human glutamatergic neurons"
Article Title: Alcohol reverses the effects of KCNJ6 (GIRK2) noncoding variants on excitability of human glutamatergic neurons
Journal: Molecular Psychiatry
doi: 10.1038/s41380-022-01818-x

Figure Legend Snippet: A Diagram outlining experimental design. Lymphocytes from subjects with or without AUD diagnosis and KCNJ6 haplotype variants were selected, reprogrammed into iPSC, induced into excitatory iNs, and analyzed by morphometry, immunocytochemistry, gene expression, and electrophysiology. B Sequencing alignment and depth analysis of bulk RNA sequencing confirmed expression of KCNJ6 mRNA in iN cultures, specifically the ENST00000609713 isoform, containing an 18.1 kilobase 3′UTR region. KCNJ6 exons are mapped to chromosome locations (marked in mb, megabases) and the position of the gene is indicated by the red box on the chromosome 21 pictogram, top, with transcription direction on the minus strand indicated by the broken arrow. Variant analysis of RNA sequences predicts a region of linkage disequilibrium of 22 SNPs, including the 3 SNPs used to select subjects (red, Table ), and 19 additional SNPs (blue, Supplementary Table ). Depth: number of sequencing reads per base aligned by position. Frequency: thickness of curved lines represents the relative frequency of splice site utilization between exons. C Single-cell RNAseq identifies a cluster of induced neurons (lower left), expressing markers consistent with neuronal function including SYP , SLC17A6 , GRIN2B , SCN3A , KCNJ3 , and KCNJ6 ; distinct from “transition neurons” that either do not express these markers or express sporadically. Additional markers are plotted in Supplementary Fig. . Isolating KCNJ6 mRNA expression, aggregated by subject and treatment, AF neurons expressed a trend towards lower levels than UN neurons ( p = 0.0508; Wald test), but treatment of 7d with IEE at 20 mM peak concentration increased AF expression above untreated ( p = 0.0225) to levels similar to UN control ( p = 0.322, not denoted on figure). D Volcano plot for untreated UN vs. AF neurons, highlighting genes significantly different (FDR > 0.05) and at least 1.5-fold changed (red dots). Genes below the fold-change cut-off are marked in blue, and those not significantly different are marked in green. Significantly different genes are listed in Supplementary Table . E Gene ontology (GO) enrichment of top 10 biological process (BP) terms for up- or downregulated genes. Plots indicate the number of regulated genes from the term and the color indicates the adjusted p value (q-value; key). All enriched terms are listed in Supplementary Tables , .
Techniques Used: Immunocytochemistry, Expressing, Sequencing, RNA Sequencing Assay, Variant Assay, Concentration Assay

Figure Legend Snippet: A Representative confocal images of GIRK2 immunoreactivity in mouse cortical neurons. Arrowheads indicate locations of GIRK2-staining puncta, with an example punctum enlarged in the inset, overlapping or adjacent to βIII-tubulin-positive processes. We observed two cellular expression patterns—one where the entire neuron is decorated with GIRK2 antibody (Supplementary Fig. ), or another where GIRK2 expression is relatively faint and observed mostly on neuronal processes, shown here. B GIRK2 expression patterns in human induced neurons (iNs), showing representative confocal images from line 420. Inset shows two adjacent puncta. GIRK2 immunoreactivity matched a pattern of process-selective expression in mouse ( A and Supplementary Fig. ), where GIRK2 was detected as relatively small (~0.5 µm diameter) puncta scattered primarily along the processes. Localization of GIRK2 immunoreactivity in human iN did not directly colocalize with synaptic vesicle marker VGLUT2 or synaptic marker Syn1 (Supplementary Fig. ), but instead was found most frequently adjacent to synapses but overlapping the shafts of the βIII tubulin-positive processes, and less so on MAP2 positive processes (Figs. 2D.a, ). Cultured neurons express βIII tubulin throughout the cell, but not as strongly in axonal processes . We previously found that processes in human iN cells stained for ankyrin G, identifying the axonal initial segment, which similarly lacked βIII-tubulin . Detection of GIRK2 primarily on βIII-tubulin + /MAP - processes, therefore, suggests pre-axonal, and likely presynaptic, localization. C Following infection of iN cultures with lentivirus expressing both KCNJ6 and mCherry, large numbers of GIRK2 + puncta are seen in representative images (line 420). D Evaluation of GIRK2 function in iNs, (a) quantification of GIRK2 expression on MAP2 + vs. βIII-tubulin + neuronal processes. GIRK2 is more abundant on βIII-tubulin processes ( p = 0.0006, one-tailed Student’s t test, n = 15 cells per group, cell line 420). (b) Basal levels (upper pie plot) of the GIRK current in iNs as percent of neurons responding with hyperpolarization to the selective GIRK activator (160 nM ML297); compared with responding percentage when GIRK2 is overexpressed (lower pie chart). (c) Representative image of iN overexpressing GIRK2, as confirmed with mCherry fluorescence. (d) Representative traces of induced action potential firing before and after GIRK activation, demonstrating the contribution of GIRK function to cell excitability. (e) Representative trace of spontaneous postsynaptic current (sEPSCs) recordings during ML297 (160 nM) GIRK activator wash-in, demonstrating a shift of 7 mV holding current (amplifier-dependent compensation of GIRK-mediated membrane hyperpolarization). (f) Quantification of neuronal excitability at baseline and following GIRK activation with 160 nM ML297 ( p = 0.015, paired Student’s t test, n = 9 cells before/after ML297, cell line 376).
Techniques Used: Staining, Expressing, Marker, Cell Culture, Infection, One-tailed Test, Fluorescence, Activation Assay

Figure Legend Snippet: A Principles of morphological analysis of induced neurons: (a) Neurite area was the total TuJ1 + (βIII-tubulin + ) staining area outside the cell soma. (b) Solidity is the area of the soma divided by its convex hull area. (c) Soma size was the area of the MAP2 + cell body. (d) Circularity compared the perimeter to the area. B Morphometry of iNs from KCNJ6 haplotype variant and affected ( AF , cyan) or unaffected ( UN , gray) individuals. Results are summed by group (left) or plotted individually by cell line (right), with subjects identified by line number (see Table —females identified with gray numbers). Individual cells are plotted as dots with the bar showing the mean, with error bars indicating the standard error of the mean (SEM). No significant differences were found in (a) soma size, (b) circularity, or (c) soma solidity, but (d) total neurite area was increased in the AF group ( p = 0.018). C Representative images of iNs from individual lines, with arrows identifying individual GIRK2 puncta (red) localized on βIII-tubulin + processes (gray). D GIRK2 expression was decreased in the AF as measured by puncta counts (a, p = 0.0012), circularity (c, p = 0.007), or solidity (d, p = 0.037), while there was no difference in puncta size (b). E Representative images of individual GIRK2 puncta (red) localized on βIII-tubulin + processes (gray). F Electrophysiological analysis of passive neuronal properties, showing no difference in (a) membrane capacitance (b) membrane resistance, or (c) spontaneous EPSCs frequency. (d) Representative sEPSCs traces for each line. (e) Spontaneous EPSCs amplitude. G Electrophysiological analysis of induced neuronal properties. (a) Quantification of current required to shift resting membrane potential to −65 mV in pA: difference by group p = 1.2 × 10 –5 ; (b) quantification of maximum number of action potentials (APs) induced with the “step” protocol, p = 0.086; (c) representative traces of APs induced with the “step” protocol; (d) quantification of number of action potentials (APs) induced with the “ramp” protocol, p = 2.0 × 10 –9 ; (e) representative traces of APs induced with the “ramp” protocol. A generalized linear model was used to evaluate group differences for morphometry and GIRK2 expression, and generalized estimation equations was used for electrophysiology results. Numbers of cell lines and replicates for each experiment are shown in Supplementary Table .
Techniques Used: Staining, Variant Assay, Expressing

Figure Legend Snippet: A Morphological analysis of IEE iNs generated from affected and unaffected individuals, showing no difference in: (a) neuronal soma size ( p = 0.32); (b) soma circularity ( p = 0.54); (c) soma solidity ( p = 0.92); and (d) total neurite area ( p = 0.98). B There was no difference in GIRK2 expression in the IEE AF group iNs compared with UN by (a) puncta counts ( p = 0.46), (b) puncta size ( p = 0.36), (c) puncta circularity ( p = 0.052), or (d) solidity ( p = 0.47). C Representative images of individual GIRK2 puncta (red) localized on βIII-Tubulin-positive processes (gray) for each cell line. D Electrophysiological analysis of passive neuronal properties in IEE iNs, showing (a) a small decrease in AF membrane capacitance ( p = 9.6 × 10 –9 ), but no difference in (b) membrane resistance ( p = 0.63), (c) spontaneous EPSCs frequency ( p = 0.32), or (d) spontaneous EPSCs amplitude ( p = 0.19). (e) Representative sEPSCs traces for each cell line. (f) The AF group exhibited no change in resting membrane potential after IEE ( p = 0.79). E Electrophysiological analysis of active neuronal properties found no difference in IEE iNs for (a) current required to shift resting membrane potential to −65 mV ( p = 0.32), (b) maximum number of action potentials (APs) induced with the “step” protocol ( p = 0.48), with (c) representative traces of APs induced with the “step” protocol, (d) number of action potentials (APs) induced with the “ramp” protocol ( p = 0.95), with (e) representative traces of APs induced with the “ramp” protocol. F Representative images from individual lines of iNs, exposed to 7d of IEE, marked with arrows pointing to individual GIRK2 puncta (red) localized on βIII-Tubulin-positive processes (gray). G Summarized results from all lines showing differences in GIRK2 expression levels before and after 7 days of 20 mM IEE with ethanol (EtOH; p = 1.0 × 10 −20 ). H Representative images of individual GIRK2 puncta (red) localized on βIII-Tubulin-positive processes (gray) prior and following 7 days 20 mM IEE with ethanol. Sample images are from line 246. I Representative images of FISH detection of KCNJ6 mRNA for each cell line. J Quantification of FISH. (a) The number of KCNJ6 puncta normalized to the number of cells in an image shows decreased expression in control AF compared with UN ( p = 8.2 × 10 −3 ) and increased expression following IEE ( p = 8.2 × 10 −3 ; using Tukey’s pairwise comparisons). (b) The percentage of KCNJ6 -expressing MAP2 + cells substantially increase in the (c) AF group but not in the (b) UN group. Numbers of KCNJ6 puncta were analyzed by expression levels per cell, as recommended by the FISH manufacturer in (d) UN or (e) AF cells. (f) KCNJ6 puncta within the neuronal soma show increases following IEE in both UN and AF groups ( p = 0.006 for genotype, Tukey’s post-hoc for UN, p = 0.01, for AF, p = 0.01). (g) A similar analysis of non-somatic puncta, presumably within neurites, showed no differences following IEE between genotypes ( p = 0.23).
Techniques Used: Generated, Expressing

Figure Legend Snippet: A Current required to shift resting membrane potential to −65 mV, in untreated (control, p = 5.9 × 10 −4 ), lentiviral KCNJ6 overexpression (over., p = 0.23), or 1 d 20 mM EtOH ( p = 0.75) cultures. B Representative traces of APs induced with the “ramp” protocol. C Quantification of maximum number of action potentials (APs) induced with “ramp” protocol, control ( p = 6.8 × 10 −6 ), overexpression ( p = 0.014), or 1 d 20 mM EtOH ( p = 0.10). D quantification of GIRK2 puncta after overexpression (line 376, one-tailed Student’s t test p = 0.04).
Techniques Used: Over Expression, One-tailed Test
girk2 (Alomone Labs)


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Girk2, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/girk2/product/Alomone Labs
Average 93 stars, based on 1 article reviews
Price from $9.99 to $1999.99
Images
1) Product Images from "Single cell transcriptomics reveals correct developmental dynamics and high-quality midbrain cell types by improved hESC differentiation"
Article Title: Single cell transcriptomics reveals correct developmental dynamics and high-quality midbrain cell types by improved hESC differentiation
Journal: bioRxiv
doi: 10.1101/2022.09.15.507987

Figure Legend Snippet: ( A and B ) Immunocytochemical staining of FOXA2 + /TH + as well as NURR1 + /TH + ( A ), as well as PITX3 + /TH + , LMO3 + /TH + , ALDH1A1 + /TH + , GIRK2 + /TH + ( B ) at day 56. Scale bar, 200 µm ( A ) and 25 µm ( B ). ( C-L ) Electrophysiological analysis of cells from days 56-73. ( C and D ) Decrease of resting membrane potential ( C ) (n = 55 cells) and reduction in input resistance ( D ) (n = 46 cells) with increasing days in vitro (DIV). ( E and F ) Percentage of cell population exhibiting the ability to generate the different spiking types in response to square current pulses as seen in example traces ( F ) (n = 12 cells for 56-59 DIV, n = 11 cells for 62-63 DIV, n = 14 cells for 64-67 DIV and n = 14 cells for 71-73 DIV). ( G ) Example trace of a spontaneous active neuron. ( H ) Percentage of each cell population spontaneously spiking (n = 13 cells for 56-59 DIV, n = 8 cells for 62-63 DIV, n = 12 cells for 64-67 DIV and n = 13 cells for 71-73 DIV). ( I ) Spontaneous action potential (AP) frequency of cells that were spontaneously spiking (n = 12 cells). ( J ) Example trace of a cell receiving two spontaneous excitatory post-synaptic currents (sEPSCs). ( K ) Cumulative distribution of sEPSC amplitudes in each cell population (Kolmogorov–Smirnov test: 56-59 DIV vs 62-67 DIV p-value = 0.243, 56-59 DIV vs 71-73 DIV p-value = 7.97 x 10 -4 , 62-67 DIV vs 71-73 DIV p-value = 9.67 x 10 -4 ). ( L and M ) HPLC analysis of whole cell dopamine content from day 28-56 ( L ) and dopamine release at day 56 ( M ). *p < 0.05, **p < 0.01 (n = 4, error bars are S.E.M.).
Techniques Used: Staining, In Vitro
rabbit k ir 3 2 (Alomone Labs)


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Rabbit K Ir 3 2, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/rabbit k ir 3 2/product/Alomone Labs
Average 93 stars, based on 1 article reviews
Price from $9.99 to $1999.99
Images
1) Product Images from "Targeting the interaction of GABA B receptors with CaMKII with an interfering peptide restores receptor expression after cerebral ischemia and inhibits progressive neuronal death in mouse brain cells and slices"
Article Title: Targeting the interaction of GABA B receptors with CaMKII with an interfering peptide restores receptor expression after cerebral ischemia and inhibits progressive neuronal death in mouse brain cells and slices
Journal: Brain Pathology
doi: 10.1111/bpa.13099

Figure Legend Snippet: R1‐Pep treatment reduced aberrant expression of K ir 3.2 channels after glutamate stress. Glutamate‐stressed neurons were treated for 16 h with R1‐Pep (10 μg/ml) or Ctrl‐Pep (10 μg/ml) and then stained with K ir 3.2 antibodies. (A) Immunofluorescence staining: top, representative images (scale bar: 5 μm); bottom, quantification of fluorescence intensities. N = 30 neurons per condition from two independent experiments. Signals were normalized to no Pep control. Two‐way ANOVA with Tukey's multiple comparison test (ns, p > 0.05; ****, p < 0.0001). (B) Western blotting: top panel, staining for K ir 3.2 channels; bottom panel, staining for total protein used for normalization. Control, untreated cultures; Glu, glutamate‐stressed cultures; Glu + R1‐Pep, glutamate‐stressed cultures treated with R1‐Pep; Glu + R1‐Pep + CGP, glutamate‐stressed cultures treated with R1‐Pep and the GABA B receptor antagonist CGP 56999. Signals were normalized to control. N = 11 cultures per condition from four independent neuron preparations. One‐way ANOVA with Tukey's multiple comparison test (ns, p > 0.05; *, p < 0.05; **, p < 0.01 ***, p < 0.0005).
Techniques Used: Expressing, Staining, Immunofluorescence, Fluorescence, Western Blot
rabbit anti girk2 (Alomone Labs)


Structured Review

Rabbit Anti Girk2, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/rabbit anti girk2/product/Alomone Labs
Average 93 stars, based on 1 article reviews
Price from $9.99 to $1999.99
Images
1) Product Images from "Alcohol reverses the effects of KCNJ6 (GIRK2) noncoding variants on excitability of human glutamatergic neurons"
Article Title: Alcohol reverses the effects of KCNJ6 (GIRK2) noncoding variants on excitability of human glutamatergic neurons
Journal: bioRxiv
doi: 10.1101/2022.05.24.493086

Figure Legend Snippet: A . Diagram outlining experimental design. Lymphocytes from subjects with or without AUD diagnosis and KCNJ6 haplotype variants were selected, reprogrammed into iPSC, induced into excitatory iNs, and analyzed by morphometry, immunocytochemistry, gene expression, and electrophysiology. B . Sequencing alignment and depth analysis of bulk RNA sequencing confirmed expression of KCNJ6 mRNA in iN cultures, specifically the ENST00000609713 isoform, containing an 18.1 kilobase 3’UTR region. Depth: number of sequencing reads per base aligned by position. Frequency: thickness of curved lines represents the relative frequency of splice site utilization between exons. Variant analysis of RNA sequences predicts a region of linkage disequilibrium of 22 SNPs, including the 3 SNPs used to select subjects (red, ), and 19 additional SNPs (blue, Supplemental Table 1). C . Single-cell RNAseq identifies a cluster of induced neurons (upper right), expressing markers consistent with neuronal function including SYP, SCN3A, SLC17A6, GRIN2B, KCNJ3 , and KCNJ6 ; distinct from “transition neurons” that either do not express these markers or express sporadically. Isolating KCNJ6 mRNA expression, aggregated by subject and treatment, AF neurons expressed a trend towards lower levels than UN neurons (p = 0.0508; Wald test), but treatment of 7d with IEE at 20 mM peak concentration increased AF expression above untreated (p = 0.0225) to levels similar to UN control (p = 0.322, not denoted on figure). D . Volcano plot for untreated UN vs AF neurons, highlighting genes significantly different (FDR > 0.05) and at least 1.5-fold changed (red dots). Genes belopuncta circularity w the fold-change cut-off are marked in green, and those not significantly different are marked in blue. Significantly different genes are listed in Supplemental Table 2. F . Gene ontology (GO) enrichment of top 10 biological process (BP) terms for up-or down-regulated genes. Plots indicate the number of regulated genes from the term and the color indicates the adjusted p-value (q-value; key). All enriched terms are listed in Supplemental Table 3.
Techniques Used: Immunocytochemistry, Expressing, Sequencing, RNA Sequencing Assay, Variant Assay, Concentration Assay

Figure Legend Snippet: A . Representative confocal images of GIRK2 immunoreactivity in mouse cortical neurons. Arrowheads indicate locations of GIRK2-staining puncta, with an example punctum enlarged in the inset, overlapping or adjacent to βIII-tubulin-positive processes. We observed two cellular expression patterns – one where the entire neuron is decorated with GIRK2 antibody (Supplemental Fig. 5), or another where GIRK2 expression is relatively faint and observed mostly on neuronal processes, shown here. B . GIRK2 expression patterns in human induced neurons (iNs), showing representative confocal images from line 420. Inset shows two adjacent puncta. C . Following infection of iN cultures with lentivirus expressing KCNJ6 and mCherry, large numbers of GIRK2 + puncta are seen in representative images (line 420). D . Evaluation of GIRK2 function in iNs, (a) quantification of GIRK2 expression on MAP2 + vs. βIII-tubulin + neuronal processes. GIRK2 is more abundant on βIII-tubulin processes (p=0.014). (b) Basal levels (upper pie plot) of the GIRK current in iNs as percent of neurons responding with hyperpolarization to the selective GIRK activator (160 nM ML297); compared with responding percentage when GIRK2 is overexpressed (lower pie chart). (c) Representative image of iN overexpressing GIRK2, as confirmed with mCherry fluorescence. (d) Representative traces of induced action potential firing before and after GIRK activation, demonstrating the contribution of GIRK function to cell excitability. (e) Representative trace of spontaneous postsynaptic potential (sEPSCs) recordings during ML297 (160 nM) GIRK activator wash-in, demonstrating a shift of 7mV holding current (amplifier-dependent compensation of GIRK-mediated membrane hyperpolarization). (f) Quantification of neuronal excitability at baseline and following GIRK activation with ML297 (160 nM). Student’s t-test was used to evaluate differences (*p < 0.05, **p < 0.01).
Techniques Used: Staining, Expressing, Infection, Fluorescence, Activation Assay

Figure Legend Snippet: A . Principles of morphological analysis of induced neurons: (a) Neurite area was the total TuJ1 + (βIII-tubulin) + staining area outside the cell soma. (b) Solidity is the area of the soma divided by its convex hull area. (c) Soma size was the area of the MAP2 + cell body. (d) Circularity compared the perimeter to the area. B . Morphometry of iNs from KCNJ6 haplotype variant and affected ( AF , cyan) or unaffected ( UN , grey) individuals. Results are summed by group (left) or plotted individually by cell line (right), with subjects identified by line number (see --females identified with grey numbers). Individual cells are plotted as dots with the bar showing the mean, with bars indicating the standard error of the mean (SEM). No significant differences were found in (a) soma size, (b) circularity, or (c) soma solidity, but total neurite area was increased in the AF group (p = 0.0007). C . Representative images of iNs from individual lines, with arrows identifying individual GIRK2 puncta (red) localized on βIII-tubulin + processes (gray). D . GIRK2 expression was decreased in the AF as measured by puncta counts (a, p = 0.043), while there was no difference in puncta size (b), circularity (c), or solidity (d). E . Representative images of individual GIRK2 puncta (red) localized on βIII-tubulin + processes (gray). F . Electrophysiological analysis of passive neuronal properties, showing no difference in (a) membrane capacitance (b) membrane resistance, or (c) spontaneous EPSCs frequency. (d) Representative sEPSCs traces for each line. (e) Spontaneous EPSCs amplitude. G . Electrophysiological analysis of active neuronal properties. (a) Quantification of current required to shift resting membrane potential to -65mV in pA: difference by group p = 0.048; (b) quantification of maximum number of action potentials (APs) induced with the “step” protocol, p = 0.014; (c) representative traces of APs induced with the “step” protocol; (d) quantification of number of action potentials (APs) induced with the “ramp” protocol, p = 0.036; (e) representative traces of APs induced with the “ramp” protocol. A generalized linear model was used to evaluate group differences for morphometry and GIRK2 expression, and generalized estimation equations was used for electrophysiology results (*p < 0.05, **p < 0.01, ***p < 0.001).
Techniques Used: Staining, Variant Assay, Expressing

Figure Legend Snippet: A . Morphological analysis of IEE iNs generated from affected and unaffected individuals, showing no difference in: (a) neuronal soma size (p = 0.38); (b) soma circularity (p = 0.47); (c) soma solidity (p = 0.87); and (d) total neurite area (p = 0.98). B . There was no difference in GIRK2 expression in the IEE AF group iNs compared with UN by (a) puncta counts (p = 0.28), (b) puncta size (p = 0.34), or (d) solidity (p = 0.43), but there was a slight increase in (c) puncta circularity (p = 0.046). C . Representative images of individual GIRK2 puncta (red) localized on βIII-Tubulin positive processes (gray) for each cell line. D . Electrophysiological analysis of passive neuronal properties in IEE iNs, showing (a) a small decrease in AF membrane capacitance (p = 9.6 × 10 −9 ), but no difference in (b) membrane resistance (p = 0.63), (c) spontaneous EPSCs frequency (p = 0.32), or (d) spontaneous EPSCs amplitude (p = 0.19). (d) Representative sEPSCs traces for each cell line. (f) The AF group exhibited no change in resting membrane potential after IEE (p = 0.79). E . Electrophysiological analysis of active neuronal properties found no difference in IEE iNs for (a) current required to shift resting membrane potential to -65mV (p = 0.32), (b) maximum number of action potentials (APs) induced with the “step” protocol (p = 0.48), with (c) representative traces of APs induced with the “step” protocol, (d) number of action potentials (APs) induced with the “ramp” protocol (p = 0.95), with (e) representative traces of APs induced with the “ramp” protocol. F . Representative images from individual lines of iNs, marked with arrows pointing to individual GIRK2 puncta (red) localized on βIII-Tubulin positive processes (gray). G . Summarized results from all lines showing differences in GIRK2 expression levels before and after 7 days of 20 mM IEE with ethanol (EtOH). H . Representative images of individual GIRK2 puncta (red) localized on βIII-Tubulin positive processes (gray) prior and following 7 days 20 mM IEE with ethanol. I . Representative images of FISH detection of KCNJ6 mRNA for each cell line. J . Quantification of FISH. (a) The number of KCNJ6 puncta normalized to the number of cells in an image shows decreased expression in control AF compared with UN (p = 1.6 × 10 −3 ), increased expression following IEE (p = 1.1 × 10 −3 ; Tukey’s pairwise comparisons for UN p = 9.0 × 10 −3 , for AF p = 5.5 × 10 −3 ). (b) The percentage of KCNJ6 -expressing MAP2 + cells substantially increase in the (c) AF group but not in the (b) UN group. Numbers of KCNJ6 puncta were analyzed by expression levels per cell, as recommended by the FISH manufacturer in (d) UN or (e) AF cells. (f) KCNJ6 puncta within the neuronal soma show increases following IEE in both UN and AF groups (p = 4.1 × 10 −4 , Tukey’s post-hoc for UN, p = 1.1 × 10 −3 , for AF, p = 1.1 × 10 −3 ). (g) A similar analysis of non-somatic puncta, presumably within neurites, showed no differences following IEE (p=0.24).
Techniques Used: Generated, Expressing

Figure Legend Snippet: A . Current required to shift resting membrane potential to -65mV, in untreated (control, p = 5.9 × 10 −4 ), lentiviral KCNJ6 overexpression (over., p = 0.23), or 1 d 20 mM IEE (EtOH, p = 0.75) cultures. B . Representative traces of APs induced with the “ramp” protocol. C . Quantification of maximum number of action potentials (APs) induced with “ramp” protocol, control (p = 6.8 × 10 −6 ), overexpression (p = 0.014), or 1 d 20 mM IEE (p = 0.10). D . quantification of GIRK2 puncta after overexpression (line 376, one-sided t-test p = 0.04).
Techniques Used: Over Expression
anti girk2 (Alomone Labs)


Structured Review

Anti Girk2, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/anti girk2/product/Alomone Labs
Average 93 stars, based on 1 article reviews
Price from $9.99 to $1999.99
Images
1) Product Images from "Functional Recovery from Human Induced Pluripotent Stem Cell-Derived Dopamine Neuron Grafts is Dependent on Neurite Outgrowth"
Article Title: Functional Recovery from Human Induced Pluripotent Stem Cell-Derived Dopamine Neuron Grafts is Dependent on Neurite Outgrowth
Journal: bioRxiv
doi: 10.1101/2022.04.19.488213

Figure Legend Snippet: Based on histological analysis of the grafts from d18 and d25 preparations, we present the total mean HuNu+cells (A), graft volume (B), the density of TH+ cells per mm 3 (C), total TH+ neurons (D), percentage of TH+ cells out of total HuNu+ cells (E), the total GIRK2+ cells (G) and the percentage of GIRK2+ cells out of TH+ cells (H). The total AADC+ cell data are depicted in (J) and the percentage of AADC+ cells relative to HuNu+ cells is in (K). Representative immunohistochemistry is presented for TH (F), GIRK2 (blue) and HuNu (brown) in (I) and AADC (brown) and HuNu (blue) in (L). Main effects of cell line or days in vitro (DIV) are stated, with p*≤0.05, p**≤0.001, error bars=±SEM.
Techniques Used: Immunohistochemistry, In Vitro
anti k ir 3 2 antibody (Alomone Labs)


Structured Review

Anti K Ir 3 2 Antibody, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/anti k ir 3 2 antibody/product/Alomone Labs
Average 93 stars, based on 1 article reviews
Price from $9.99 to $1999.99
Images
1) Product Images from "Targeting the Interaction of GABA B Receptors With CHOP After an Ischemic Insult Restores Receptor Expression and Inhibits Progressive Neuronal Death"
Article Title: Targeting the Interaction of GABA B Receptors With CHOP After an Ischemic Insult Restores Receptor Expression and Inhibits Progressive Neuronal Death
Journal: Frontiers in Pharmacology
doi: 10.3389/fphar.2022.870861

Figure Legend Snippet: R2-Pep normalize OGD-mediated dysregulation of K ir 3.2 channels. Cultures were stressed for 1 h with OGD and immediately treated with R2-Pep, Ctrl-Pep or remained untreated. After 16 h neurons were immunostained for K ir 3.2 channel expression. Top: representative images (scale bar: 5 μm). Bottom: quantification of fluorescence intensities. The fluorescence intensity of the untreated neurons served as control. The data represent the mean ± S.E.M. of 37–38 neurons from two independent preparations. ns: p > 0.05; ****, p < 0.0001; two-way ANOVA with Tukey’s multiple comparisons test.
Techniques Used: Expressing, Fluorescence