Journal: PLoS ONE

Article Title: Targeted disruption of glycogen synthase kinase-3β in cardiomyocytes attenuates cardiac parasympathetic dysfunction in type 1 diabetic Akita mice

doi: 10.1371/journal.pone.0215213

Figure Lengend Snippet: Levels of SREBP-1 and GIRK1/4 in atria from placebo and tamoxifen treated GSK3β fl/fl/ Cre + Akita mice determined by Western blot analysis of atrial extracts of 6 mice in each group. B : Densitometry analysis of SREBP-1 and GIRK1/4 from A . Data were normalized to the expression of GAPDH. ** P <0.01 compared to control Akita mice.

Article Snippet: The GIRK4 specific antibody was from Santa Cruz Biotechnology and the GIRK1 specific antibody was from Alomone Labs (Israel).

Techniques: Western Blot, Analysis, Expressing, Control

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: bioRxiv

Article Title: Decoding subtype development and function in human pluripotent stem cell-derived midbrain dopaminergic neurons

doi: 10.64898/2026.09.23.753784

Figure Lengend Snippet: a , Experimental schematic of TGFβ pathway modulation during mDA progenitor patterning. b, PITX3::GFP reporter design (top), flow cytometry of PITX3 CRISPRa activation (middle), and immunofluorescence (IF) for PITX3 and TH at D30 (bottom). c, Flow cytometry of PITX3, CD171 and CD99 at D25. d, Flow cytometry of ALDH activity and CD99 at D25. e, Flow cytometry of PITX3 expression and ALDH activity in the indicated gated subpopulations, with quantification of the percentage of PITX3+ALDH+ cells in each subpopulation. One-way ANOVA; n = 4. f, RT–qPCR of the indicated genes in D25 control and Activin A/BMP7-patterned populations. Student’s t -test; n = 3. g, Flow cytometry of PITX3 and ALDH activity in the indicated CD99-CD56+ populations. h, IF for PITX3, ALDH1A1 and CALB1 at D26. Scale bars, 100 μm. i, Schematic illustrating mDA subtype patterning through modulation of TGF-β signaling. j, Schematic of the day 25 sorting strategy and RT–qPCR analysis of the indicated genes in sorted day 25 populations. One-way ANOVA; n = 3–4. k, IF for ALDH1A1, PITX3 and CALB1 at D26 in populations derived from D25 sorted cells (left), with quantification of relative ALDH1A1 expression (right). Scale bars, 100 μm. One-way ANOVA; n = 4–8. l, Flow cytometry of TH and ALDH activity in the indicated CD99− and CD99+ populations, with quantification of the percentage of CD99-TH+ALDH+ cells. One-way ANOVA; n = 3. m . Schematic of hPSC-A9/A10 transplantation into the mouse striatum (top); IF staining for GIRK2, TH and CALB1 in A9 and A10 grafts at 6 months post-transplantation, with quantification of GIRK2+TH+ and CALB1+TH+ cells in the grafts. Student’s t -test; n = 3. All error bars in represent s.e.m.

Article Snippet: Sections were incubated overnight at 4 °C with primary antibodies against GIRK2 (1:100; Alomone Labs, APC_006-GP) and calbindin (1:200; Cell Signaling Technology, 13176).

Techniques: Flow Cytometry, Activation Assay, Immunofluorescence, Activity Assay, Expressing, Quantitative RT-PCR, Control, Analysis, Derivative Assay, Transplantation Assay, Staining, FACS