Journal: Scientific Reports

Article Title: Mutual action by Gγ and Gβ for optimal activation of GIRK channels in a channel subunit-specific manner

doi: 10.1038/s41598-018-36833-y

Figure Lengend Snippet: Distal C-terminus of GIRK1 is important for Gγ activation of GIRK1* and GIRK1/3. ( A–C ) Deletion of dCT abolishes the activating effect of YFP-Gγ (2.5 ng RNA) on GIRK1*. ( A , B ) show representative traces of GIRK1* ( A ) and GIRK1* Δ121 ( B ) expressed alone or with YFP-Gγ. m2R was expressed in all cases. ( C ) Summary of the experiment. Numbers above data sets are n, N. Statistics: t-test for each pair with and without YFP-Gγ. ***p < 0.001, N.S., not statistically significant. ( D – F ) Expression of Gγ tandem (Gγ Tan) does not affect the expression of GIRK1/3 in the PM ( D,E ) and activates GIRK1/3 but not GIRK1 Δ121 /3 (GIRK1 Δ121 /3; F ). Oocytes were stained with an antibody against GIRK3. Representative images of giant excised PM patches are shown in ( D ) and summary of measurements in ( E ) Gγ tandem did not affect the expression of GIRK1/3, but reduced the expression of GIRK1 Δ121 /3 when expressed at a high dose (p < 0.05 for 2 ng Gγ tandem). A.U., arbitrary units. n = 12–16 membranes in each group, N = 1. ( F ) Summary of the effects of Gγ tandem and of Gβγ on I basal . n = 10–15 cells in each group, N = 2. Gγ tandem and Gβγ significantly increased I basal of GIRK1/3 (***p ≤ 0.001 vs. GIRK1/3 alone). GIRK1 Δ121 /3 was not affected by Gγ tandem but was activated by Gβγ (###p < 0.001 vs. GIRK1 Δ121 /3 alone).

Article Snippet: Anti-Kir3.1 (GIRK1) antibody (Alomone labs, APC-005) or Anti-Kir3.3 (GIRK3) antibody (Alomone labs, APC-038) were applied at 1:200 or 1:100 dilution respectively, for 45 minutes at 37 °C.

Techniques: Activation Assay, Expressing, Staining

Journal: Scientific Reports

Article Title: Mutual action by Gγ and Gβ for optimal activation of GIRK channels in a channel subunit-specific manner

doi: 10.1038/s41598-018-36833-y

Figure Lengend Snippet: List of cDNA constructs.

Article Snippet: Anti-Kir3.1 (GIRK1) antibody (Alomone labs, APC-005) or Anti-Kir3.3 (GIRK3) antibody (Alomone labs, APC-038) were applied at 1:200 or 1:100 dilution respectively, for 45 minutes at 37 °C.

Techniques: Construct, Mutagenesis, Expressing, Sequencing, Ligation

Journal: Developmental biology

Article Title: Genetic Dissection of Midbrain Dopamine Neuron Development in vivo

doi: 10.1016/j.ydbio.2012.09.019

Figure Lengend Snippet: Quantification of cells in control and mutant mice reveals significant differences in the average number of TH+ and double positive neurons in the VTA and SNc. The average number of TH+ neurons and the 95% confidence intervals of counts from the VTA (A, B) and SNc (C, D) of control (dark grey shaded bars) and mutant mice (dark brown bars) in ventral, intermediate and dorsal planes. (A) The average number of TH+/Calb+ neurons in the VTA of mutant mice (light brown bars) was significantly lower than that of control mice (light grey bars) in all planes quantified. Calb+ neurons in the VTA indicated that the medial domain was not devoid of cells. (B) There were no double immunolabeled TH+/GIR2K+ neurons observed in the ventral VTA of any of the mutant GIRK2-stained slices consistent with the loss of VTA. (C) Although TH+ neurons were diminished in SNc, the average number of TH+/Calb+ neurons in the SNc was not significantly different between controls (light grey bars) and mutants (light brown bars) at any of the three horizontal planes. The blue arrowheads points to the average of TH+/Calb+ neurons. (D) The average number of TH+/GIRK2+ neurons in the SNc of mutant mice was significantly lower than that of control mice in all planes quantified. Images in A, C display TH+ (green), Calb+ (red) and TH+/Calb+ (yellow) cells while images in B, D display TH+ (green), GIRK2+ (red) and TH+/GIRK2+ (yellow) cells. Gray shading of bars indicates data from control animals with dark gray bars corresponding to average TH+ cell counts and nested light gray bars corresponding to average double positive cell counts. Brown shading indicates data from mutant animals with dark brown bars corresponding to average TH+ cell counts and nested lighter brown bars corresponding to average double positive cell counts.

Article Snippet: Calb indin (CALB) and g -protein i nward r ectifying potassium ( K ) channel 2 (GIRK2) were detected with anti-CALB (Swant; 1:1000) or anti-Girk2 (Alomone labs; 1:80), respectively.

Techniques: Mutagenesis, Immunolabeling, Staining