trpv3  (Alomone Labs)

Journal: The Journal of investigative dermatology

Article Title: PAR2 mediates itch via TRPV3 signaling in keratinocytes

doi: 10.1016/j.jid.2020.01.012

Figure Lengend Snippet: TRPV3 is involved in PAR2/Ca 2+ signaling in keratinocytes. ( a ) The percentages of responding keratinocytes from WT and Trpv3 −/− mice stimulated with histamine (100 μM) and SLIGRL (50 μM). ***p

Article Snippet: Primary antibodies used for immunofluorescence of cultured primary keratinocytes were: anti-TRPV3 antibody (Alomone Labs, 1:300) and anti-PAR2 antibody (Santa Cruz, 1:100).

Techniques: Mouse Assay

Journal: The Journal of investigative dermatology

Article Title: PAR2 mediates itch via TRPV3 signaling in keratinocytes

doi: 10.1016/j.jid.2020.01.012

Figure Lengend Snippet: Topical blockade of PAR2 and TRPV3 attenuated the manifestations of AD mouse model. ( a-d ) Ear appearance of WT mice topically applied with vehicle as a control ( a ), MC-903 ( b ), MC-903+74a ( c ), and MC-903+FSLLRY ( d ) for 7 days. ( e-h ) Hematoxylin-eosin staining showing changes in hyperkeratosis and inflammatory infiltration in the ears of corresponding mice in panels a-d , respectively. Scale = 50 μm. ( i. j ) The scratching numbers ( i ) and ear thickness increment ( j ) of mice treated with MC-903, MC-903+74a and MC-903+FSLLRY for 7 days. One-way ANOVA followed by Tukey’s post hoc . n = 6–8 mice per group. ( k ) Diagram illustrating PAR2-TRPV3 signaling pathway in itch.

Article Snippet: Primary antibodies used for immunofluorescence of cultured primary keratinocytes were: anti-TRPV3 antibody (Alomone Labs, 1:300) and anti-PAR2 antibody (Santa Cruz, 1:100).

Techniques: Mouse Assay, Staining

Journal: The Journal of investigative dermatology

Article Title: PAR2 mediates itch via TRPV3 signaling in keratinocytes

doi: 10.1016/j.jid.2020.01.012

Figure Lengend Snippet: TRPV3 is involved in PAR2-induced acute itch. ( a ) Acute itch behaviors of Trpv3 −/− mice and their WT littermates induced by intradermal injections of SLIGRL (50 μg), trypsin (100 μg), 2fly (10 μg), histamine (200 μg), and BAM 8-22 (150 μg). Unpaired t tests. n = 6–8. ( b ) WT mice were pre-injected with vehicle (Veh) (8% DMSO), 74a (150 μg, i.d.) or FSLLRY (100 μg, i.d.) 15 min before testing the scratching behaviors elicited by SLIGRL, trypsin, or 2fly. One-way ANOVA followed by Tukey’s post hoc . n = 6-8 mice per group. ( c ) TRPV3 expression in the skin and DRG of Trpv3 +/+ and Trpv3 −/− mice as determined by western blot. ( d ) Double immunofluorescence staining of PAR2 (green) and TRPV3 (red) in mouse keratinocytes. Scale bar, 20 μm. ( e ) Venn diagram showing the overlapping of PAR2 and TRPV3 in mouse keratinocytes. *p

Article Snippet: Primary antibodies used for immunofluorescence of cultured primary keratinocytes were: anti-TRPV3 antibody (Alomone Labs, 1:300) and anti-PAR2 antibody (Santa Cruz, 1:100).

Techniques: Mouse Assay, Injection, Expressing, Western Blot, Double Immunofluorescence Staining

Journal: The Journal of investigative dermatology

Article Title: PAR2 mediates itch via TRPV3 signaling in keratinocytes

doi: 10.1016/j.jid.2020.01.012

Figure Lengend Snippet: The G-protein/PLC/Ca 2+ pathway mediated PAR2 activation in keratinocytes, and PAR2 and TRPV3 were necessary for adequate TSLP release from keratinocytes. ( a-c ) Representative traces showing that Ca 2+ transients elicited by SLIGRL (50 μM) in WT mouse keratinocytes were completely inhibited by co-incubation with 100 μM gallein, a G βγ -protein inhibitor ( a ) and 10 μM U73122, a PLC inhibitor ( b ), but were partially inhibited by co-incubation with 10 μM DBHQ, an ATPase inhibitor ( c ). ( d ) Quantified data showing the percentages of SLIGRL-responsive keratinocytes before and after the incubation of gallein, U73122 or DBHQ. *p

Article Snippet: Primary antibodies used for immunofluorescence of cultured primary keratinocytes were: anti-TRPV3 antibody (Alomone Labs, 1:300) and anti-PAR2 antibody (Santa Cruz, 1:100).

Techniques: Planar Chromatography, Activation Assay, Incubation

Journal: The Journal of investigative dermatology

Article Title: PAR2 mediates itch via TRPV3 signaling in keratinocytes

doi: 10.1016/j.jid.2020.01.012

Figure Lengend Snippet: PAR2 and TRPV3 were involved in the pathogenesis of chronic itch. ( a ) WT, Trpv3 −/− and Par2 −/− mice were treated daily with a topical application of MC-903. Scratching numbers were recorded on day 1, 3, 5, and 7. Two-way ANOVA followed by Bonferroni post hoc . n = 6–8 mice per group. ( b ) Real time RT-PCR showing the relative levels of TRPV3 and PAR2 mRNA in pruritic skin lesions of AD patients and normal control. Unpaired t test. n = 12. ( c, d ) Relative levels of PAR2 mRNA ( c ) and TRPV3 mRNA ( e ) in the ears of MC-903-treated mice on day 5. Two-way ANOVA with Bonferroni post hoc . n = 4–5. Data are presented as mean ± s.e.m. *p

Article Snippet: Primary antibodies used for immunofluorescence of cultured primary keratinocytes were: anti-TRPV3 antibody (Alomone Labs, 1:300) and anti-PAR2 antibody (Santa Cruz, 1:100).

Techniques: Mouse Assay, Quantitative RT-PCR

Journal: Journal of Cellular and Molecular Medicine

Article Title: The effect of Sirt1 deficiency on Ca2+ and Na+ regulation in mouse ventricular myocytes, et al. The effect of Sirt1 deficiency on Ca2+ and Na+ regulation in mouse ventricular myocytes

doi: 10.1111/jcmm.15327

Figure Lengend Snippet: Ca 2+ regulatory proteins in control and Sirt1 −/− ventricular myocytes. A, Representative immunoblot and mean data for Cav1.2, SERCA2a, NCX and CaMKII in control and Sirt1 −/− mice ventricular myocytes. B, Normalized densitometry for the protein levels (α‐Tubulin was used as an internal control; Control N = 6 and Sirt1 −/− N = 6; * P

Article Snippet: The cardiac protein extracts were separated by sodium dodecyl sulphate polyacrylamide gel electrophoresis and then transferred to polyvinylidene difluoride membranes (Millipore) that were incubated with the listed antibodies: Cav1.2 (1:1000, AACC‐033, rabbit polyclonal antibody; Alomone Labs, Jerusalem, Israel), CaMKII (1:1000, sc‐5306, mouse monoclonal antibody; Santa Cruz Biotechnology, Dallas, USA), NCX (1:1000, mouse monoclonal antibody, ab2869; Abcam), SERCA2a (1:5000, sc‐376235, mouse monoclonal antibody; Santa Cruz Biotechnology, Dallas, USA) and α‐tubulin (1:10 000, sc‐5286, mouse monoclonal antibody; Santa Cruz Biotechnology, Dallas, USA).

Techniques: Mouse Assay