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ym26734  (Tocris)
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Figure 6. Piezo1 mediates TRPV4 channel opening in pancreatic stellate cells. (A) Immunostaining of GFAP and TRPV4 in human PSCs. (B) TRPV4 agonist GSK101 (100 nM) effects on [Ca2+]i in human PSCs with and without the TRPV4 blocker HC067 (1 μM). (C) GSK101 (100 nM) effects on [Ca2+]i in PSCs from 3 bio- logical samples were blocked with the TRPV4 antagonist HC067 (1 μM) (from 18 to 37 cells). (D) Immunostaining of GFAP and TRPV4 in mouse PSCs. (E and F) Traces and graph represent the effects of the TRPV4 agonist GSK101 (100 nM) on [Ca2+]i in mouse PSCs with and without the TRPV4 blocker HC067 (1 μM) (from 18 cells). (G–I) Effects of Yoda1 (25 μM) on [Ca2+]i in PSCs from WT and TRPV4-KO mice. (I) Statistical analyses of peak and sustained [Ca2+]i elevation (from 24 to 32 cells). The sustained [Ca2+]i elevation was measured at 6 minutes after Yoda1. (J and K) Effects of phospholipase A2 blockers AACOCF3 (30 μM) and <t>YM26734</t> (10 μM) on Yoda1-induced (25 μM) [Ca2+]i in PSCs (from 24 to 26 cells). The sustained calcium rise was measured at 8 minutes after Yoda1 application. (L–N) Fluid shear stress (12 dyne/cm2) was applied for 1 minute in PSCs from WT and TRPV4-KO mice and TRPV4-KO mice with GsMTx4 (5 μM). In panels G, H, and L–N, each colored line represents the response of a single cell. (O) Quantification of peak [Ca2+]i following shear stress (12 dyne/cm2) for 1 minute in TRPV4-KO PSCs with and without GsMTx4 (from 24 cells). Statistical analyses were calculated using 2-tailed Student’s t test. *P ≤ 0.05 and ****P ≤ 0.0001. Scale bar: 10 μm.
Ym26734, supplied by Tocris, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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compound 54 (ym-26734)  (Yamanouchi Pharmaceutical Co)
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Yamanouchi Pharmaceutical Co compound 54 (ym-26734)
Figure 6. Piezo1 mediates TRPV4 channel opening in pancreatic stellate cells. (A) Immunostaining of GFAP and TRPV4 in human PSCs. (B) TRPV4 agonist GSK101 (100 nM) effects on [Ca2+]i in human PSCs with and without the TRPV4 blocker HC067 (1 μM). (C) GSK101 (100 nM) effects on [Ca2+]i in PSCs from 3 bio- logical samples were blocked with the TRPV4 antagonist HC067 (1 μM) (from 18 to 37 cells). (D) Immunostaining of GFAP and TRPV4 in mouse PSCs. (E and F) Traces and graph represent the effects of the TRPV4 agonist GSK101 (100 nM) on [Ca2+]i in mouse PSCs with and without the TRPV4 blocker HC067 (1 μM) (from 18 cells). (G–I) Effects of Yoda1 (25 μM) on [Ca2+]i in PSCs from WT and TRPV4-KO mice. (I) Statistical analyses of peak and sustained [Ca2+]i elevation (from 24 to 32 cells). The sustained [Ca2+]i elevation was measured at 6 minutes after Yoda1. (J and K) Effects of phospholipase A2 blockers AACOCF3 (30 μM) and <t>YM26734</t> (10 μM) on Yoda1-induced (25 μM) [Ca2+]i in PSCs (from 24 to 26 cells). The sustained calcium rise was measured at 8 minutes after Yoda1 application. (L–N) Fluid shear stress (12 dyne/cm2) was applied for 1 minute in PSCs from WT and TRPV4-KO mice and TRPV4-KO mice with GsMTx4 (5 μM). In panels G, H, and L–N, each colored line represents the response of a single cell. (O) Quantification of peak [Ca2+]i following shear stress (12 dyne/cm2) for 1 minute in TRPV4-KO PSCs with and without GsMTx4 (from 24 cells). Statistical analyses were calculated using 2-tailed Student’s t test. *P ≤ 0.05 and ****P ≤ 0.0001. Scale bar: 10 μm.
Compound 54 (Ym 26734), supplied by Yamanouchi Pharmaceutical Co, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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ym 26734  (Cayman Chemical)
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Cayman Chemical ym 26734
Figure 6. Piezo1 mediates TRPV4 channel opening in pancreatic stellate cells. (A) Immunostaining of GFAP and TRPV4 in human PSCs. (B) TRPV4 agonist GSK101 (100 nM) effects on [Ca2+]i in human PSCs with and without the TRPV4 blocker HC067 (1 μM). (C) GSK101 (100 nM) effects on [Ca2+]i in PSCs from 3 bio- logical samples were blocked with the TRPV4 antagonist HC067 (1 μM) (from 18 to 37 cells). (D) Immunostaining of GFAP and TRPV4 in mouse PSCs. (E and F) Traces and graph represent the effects of the TRPV4 agonist GSK101 (100 nM) on [Ca2+]i in mouse PSCs with and without the TRPV4 blocker HC067 (1 μM) (from 18 cells). (G–I) Effects of Yoda1 (25 μM) on [Ca2+]i in PSCs from WT and TRPV4-KO mice. (I) Statistical analyses of peak and sustained [Ca2+]i elevation (from 24 to 32 cells). The sustained [Ca2+]i elevation was measured at 6 minutes after Yoda1. (J and K) Effects of phospholipase A2 blockers AACOCF3 (30 μM) and <t>YM26734</t> (10 μM) on Yoda1-induced (25 μM) [Ca2+]i in PSCs (from 24 to 26 cells). The sustained calcium rise was measured at 8 minutes after Yoda1 application. (L–N) Fluid shear stress (12 dyne/cm2) was applied for 1 minute in PSCs from WT and TRPV4-KO mice and TRPV4-KO mice with GsMTx4 (5 μM). In panels G, H, and L–N, each colored line represents the response of a single cell. (O) Quantification of peak [Ca2+]i following shear stress (12 dyne/cm2) for 1 minute in TRPV4-KO PSCs with and without GsMTx4 (from 24 cells). Statistical analyses were calculated using 2-tailed Student’s t test. *P ≤ 0.05 and ****P ≤ 0.0001. Scale bar: 10 μm.
Ym 26734, supplied by Cayman Chemical, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Figure 6. Piezo1 mediates TRPV4 channel opening in pancreatic stellate cells. (A) Immunostaining of GFAP and TRPV4 in human PSCs. (B) TRPV4 agonist GSK101 (100 nM) effects on [Ca2+]i in human PSCs with and without the TRPV4 blocker HC067 (1 μM). (C) GSK101 (100 nM) effects on [Ca2+]i in PSCs from 3 bio- logical samples were blocked with the TRPV4 antagonist HC067 (1 μM) (from 18 to 37 cells). (D) Immunostaining of GFAP and TRPV4 in mouse PSCs. (E and F) Traces and graph represent the effects of the TRPV4 agonist GSK101 (100 nM) on [Ca2+]i in mouse PSCs with and without the TRPV4 blocker HC067 (1 μM) (from 18 cells). (G–I) Effects of Yoda1 (25 μM) on [Ca2+]i in PSCs from WT and TRPV4-KO mice. (I) Statistical analyses of peak and sustained [Ca2+]i elevation (from 24 to 32 cells). The sustained [Ca2+]i elevation was measured at 6 minutes after Yoda1. (J and K) Effects of phospholipase A2 blockers AACOCF3 (30 μM) and YM26734 (10 μM) on Yoda1-induced (25 μM) [Ca2+]i in PSCs (from 24 to 26 cells). The sustained calcium rise was measured at 8 minutes after Yoda1 application. (L–N) Fluid shear stress (12 dyne/cm2) was applied for 1 minute in PSCs from WT and TRPV4-KO mice and TRPV4-KO mice with GsMTx4 (5 μM). In panels G, H, and L–N, each colored line represents the response of a single cell. (O) Quantification of peak [Ca2+]i following shear stress (12 dyne/cm2) for 1 minute in TRPV4-KO PSCs with and without GsMTx4 (from 24 cells). Statistical analyses were calculated using 2-tailed Student’s t test. *P ≤ 0.05 and ****P ≤ 0.0001. Scale bar: 10 μm.

Journal: JCI insight

Article Title: Piezo1-mediated stellate cell activation causes pressure-induced pancreatic fibrosis in mice.

doi: 10.1172/jci.insight.158288

Figure Lengend Snippet: Figure 6. Piezo1 mediates TRPV4 channel opening in pancreatic stellate cells. (A) Immunostaining of GFAP and TRPV4 in human PSCs. (B) TRPV4 agonist GSK101 (100 nM) effects on [Ca2+]i in human PSCs with and without the TRPV4 blocker HC067 (1 μM). (C) GSK101 (100 nM) effects on [Ca2+]i in PSCs from 3 bio- logical samples were blocked with the TRPV4 antagonist HC067 (1 μM) (from 18 to 37 cells). (D) Immunostaining of GFAP and TRPV4 in mouse PSCs. (E and F) Traces and graph represent the effects of the TRPV4 agonist GSK101 (100 nM) on [Ca2+]i in mouse PSCs with and without the TRPV4 blocker HC067 (1 μM) (from 18 cells). (G–I) Effects of Yoda1 (25 μM) on [Ca2+]i in PSCs from WT and TRPV4-KO mice. (I) Statistical analyses of peak and sustained [Ca2+]i elevation (from 24 to 32 cells). The sustained [Ca2+]i elevation was measured at 6 minutes after Yoda1. (J and K) Effects of phospholipase A2 blockers AACOCF3 (30 μM) and YM26734 (10 μM) on Yoda1-induced (25 μM) [Ca2+]i in PSCs (from 24 to 26 cells). The sustained calcium rise was measured at 8 minutes after Yoda1 application. (L–N) Fluid shear stress (12 dyne/cm2) was applied for 1 minute in PSCs from WT and TRPV4-KO mice and TRPV4-KO mice with GsMTx4 (5 μM). In panels G, H, and L–N, each colored line represents the response of a single cell. (O) Quantification of peak [Ca2+]i following shear stress (12 dyne/cm2) for 1 minute in TRPV4-KO PSCs with and without GsMTx4 (from 24 cells). Statistical analyses were calculated using 2-tailed Student’s t test. *P ≤ 0.05 and ****P ≤ 0.0001. Scale bar: 10 μm.

Article Snippet: The chemicals used in calcium imaging experiments included: GsMTx4 (Abcam, catalog ab141871), Yoda1 (Tocris, catalog 5586), GSK1016790A (MilliporeSigma, catalog G0798), HC067047 (Tocris, catalog 4100), YM26734 (Tocris, catalog 2522), and AACOCF3 (Tocris, catalog 1462).

Techniques: Immunostaining, Shear