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u73122  (Tocris)
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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 <t>U73122</t> 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
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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 <t>U73122</t> 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
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plc inhibitor u73122  (Tocris)
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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 <t>U73122</t> 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
Plc Inhibitor U73122, supplied by Tocris, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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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 <t>U73122</t> 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
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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 <t>U73122</t> 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
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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

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