Journal: Journal of Cell Science

Article Title: Molecular insights into profilin 1-dependent regulation of cellular phosphatidylinositol (4,5)-bisphosphate

doi: 10.1242/jcs.265025

Figure Lengend Snippet: PIP 2 attenuation upon loss of Pfn1 is reversible by PLC inhibition. (A,B) Representative PIP 2 staining images (A; 40×) of control and Pfn1 KO MDA 231 cells, treated with either inactive ( U73343 ) or active (U73122) pan-PLC inhibitor drug (magnified images of PIP 2 staining of selected regions of interest are shown alongside). (B) The associated PM PIP 2 quantification for the various groups (all data normalized to the average readout of control cells treated with the U73343 compound). A total of ∼450 cells/group were analyzed from three independent experiments. Scale bar: 40 μm. (C–E) Representative PLCβ3 and Pfn1 immunoblots of lysates of MDA-231 cells following transient transfection of the indicated siRNAs (non-targeting control siRNA transfection served as control); GAPDH blot serves as the loading control (C). Representative PIP 2 staining images fluorescence images (40×) of MDA-231 cells transfected with the indicated siRNAs (magnified images of PIP 2 staining of selected regions of interest of each group are shown underneath) (D). Scale bar: 80 μm. (E) Associated quantification to compare the PM PIP 2 intensity between the various groups of cells based on the results of three independent experiments. In B,E, the box represents the data falling within the 25th to 75th percentile range, and a line denoting the median value of the data set. Whiskers represent 1.5× the interquartile range above and below the 25th and 75th percentiles. * P <0.05; *** P <0.001 (one-way ANOVA with Tukey HSD post-hoc test for multiple comparisons).

Article Snippet: In experiments involving pan-PLC inhibitor U73122 and its inactive control U73343 (Medchem Express, # HY108630 ), the dry compounds were diluted in DMSO to a stock concentration of 10 mM and added to cells at a final concentration of 2 μM after 24 h of serum stimulation.

Techniques: Inhibition, Staining, Control, Western Blot, Transfection, Fluorescence

Journal: Scientific Reports

Article Title: Beta2-adrenergic receptor activation increases susceptibility to arrhythmogenesis in aging heart by impairing repolarization reserve via G i βγ–PLC signaling pathway

doi: 10.1038/s41598-026-54675-x

Figure Lengend Snippet: Involvement of G i βγ–PLC signaling in ventricular arrhythmias in isolated perfused hearts. (A, C, E, G) Representative ECG waveforms from ex vivo aging guinea pig hearts pretreated with ML277 (1 µM, A), PTX (1.5 µg/mL, C), gallein (100 µM, E) or U73122 (1 µM, G), followed by SAL (5 and 10 µM). (B, D, F, H) Percentage change in QTc interval relative to baseline after SAL treatment in hearts pretreated with ML277 (B), PTX (D), gallein (F), or U73122 (H). n = 4 hearts per group. Data are expressed as mean ± SEM. * P < 0.05, ** P < 0.01 vs. the pretreatment condition.

Article Snippet: U73122 (MedChemExpress, USA) was also prepared as a stock solution in DMSO.

Techniques: Isolation, Ex Vivo

Journal: Scientific Reports

Article Title: Beta2-adrenergic receptor activation increases susceptibility to arrhythmogenesis in aging heart by impairing repolarization reserve via G i βγ–PLC signaling pathway

doi: 10.1038/s41598-026-54675-x

Figure Lengend Snippet: Effects of G i βγ–PLC signaling inhibition on IP 3 levels in myocardial homogenates from aged hearts. ( A ) Standard curve for the IP 3 ELISA. Serial dilutions of IP 3 standards (0–10,000 pg/mL) used to generate a four-parameter logistic (4-PL) curve R 2 = 0.996. ( B ) IP 3 content in myocardial tissue after treatment with SAL (10 µM) alone or in combination with PTX (1.5 µg/mL), gallein (100 µM), or U73122 (1 µM). ( C ) Fold change in IP 3 levels normalized to the control condition. n = 5 hearts per group. Data are expressed as mean ± SEM. * P < 0.05, ** P < 0.01 vs. the contorl condition.

Article Snippet: U73122 (MedChemExpress, USA) was also prepared as a stock solution in DMSO.

Techniques: Inhibition, Enzyme-linked Immunosorbent Assay, Control

Journal: Scientific Reports

Article Title: Beta2-adrenergic receptor activation increases susceptibility to arrhythmogenesis in aging heart by impairing repolarization reserve via G i βγ–PLC signaling pathway

doi: 10.1038/s41598-026-54675-x

Figure Lengend Snippet: Involvement of G i βγ–PLC signaling in cell arrhythmias in hiPSC-CMs. (A1–A5) Representative extracellular field potential (EFP) waveforms recorded from control hiPSC-CMs under baseline conditions or after pretreatment with ML277 (1 µM, A2), PTX (1.5 µg/mL, A3), gallein (100 µM, A4) and U73122 (1 µM, A5), followed by salbutamol (SAL, 10 µM). (A6) Normalized FPDc from recordings shown in A1–A5. (B1–B5) Representative EFP waveforms from D-gal–induced aging hiPSC-CMs under the same experimental conditions. (B6) Normalized FPDc from recordings shown in B1–B5. (C) Percentage change in FPDc from recordings shown in A and B. (D) Incidence of tachyarrhythmias induced by SAL. (E) Representative types of cellular arrhythmia recorded in hiPSC-CMs. (F) Heatmap of arrhythmia scores under different treatment conditions. n ≥ 3 per group. Data are expressed as mean ± SEM. ** P < 0.01 vs. control within the same group. # P < 0.05 for intergroup comparisons.

Article Snippet: U73122 (MedChemExpress, USA) was also prepared as a stock solution in DMSO.

Techniques: Control

Journal: Journal of Biomedical Science

Article Title: OPRM1/MRGPRX1 heterodimers drive opioid-induced itch through a peripheral mechanism

doi: 10.1186/s12929-026-01238-x

Figure Lengend Snippet: DAMGO-induced calcium mobilization in OPRM1/MRGPRX1 is mediated by Gα q/11 and phospholipase C. A Schematic of inhibition assay. YM254890, a Gα q/11 inhibitor, is applied prior to DAMGO to assess Gα q/11 involvement. B Representative pseudocolor images of HEK293T cells co-expressing hOPRM1 and hMRGPRX1 (hOPRM1/hMRGPRX1) before (top) and after (bottom) treatment with 10 µM DAMGO. C Pretreatment with 10 µM YM254890, a selective Gα q/11 inhibitor, significantly reduced DAMGO-induced calcium mobilization. Scale bars indicate 50 µm. D Schematic of inhibitor assay for Gα i/o and PLC contributions. Pertussis toxin (PTX) blocks Gα i/o , whereas U73122 inhibits PLC. E Time-course of calcium responses (F/F 0 ) in hOPRM1/hMRGPRX1 cells treated with DAMGO under various conditions: control, PTX (500 ng/mL, Gα i/o inhibitor), U73122 (10 µM, PLC inhibitor), and YM254890 (10 µM, Gα q/11 inhibitor). F Quantification of peak responses (ΔPeak F/F₀) for the conditions in ( D ). Numbers shown inside the bars indicate the sample sizes (n). Statistical significance is indicated as follows: ns: not significant, *p < 0.05

Article Snippet: The following reagents were purchased from Sigma-Aldrich (Seoul, Korea): [D-Ala 2 , N-Me-Phe 4 , Gly 5 -ol]-Enkephalin (DAMGO), Leucine-Enkephalin (Leu-Enk), Naltrexone, Berbamine, U73122, Pertussis toxin (PTX), MC903 (also known as calcipotriol), Compound 48/80 (C48/80), Bovine adrenal medulla 8–22 (BAM8–22), QWF, and Forskolin. β-endorphin, Endomorphin-1, Endomorphin-2, [D-Pen 2 , D-Pen 5 ]-Enkephalin (DPDPE) were purchased from Tocris Bioscience (Bristol, UK).

Techniques: Inhibition, Expressing, Control