Journal: British Journal of Pharmacology

Article Title: Functional expression and pharmacological modulation of TRPM3 in human sensory neurons

doi: 10.1111/bph.14994

Figure Lengend Snippet: TaqMan™ gene expression assays used in this study

Article Snippet: Sample threshold cycles were analysed relative to housekeeping gene HPRT as endogenous control (ΔC t ), and data are shown as 2 −ΔCt (mean ± SEM). table ft1 table-wrap mode="anchored" t5 Table 1 caption a7 Gene Assay TRPA1 Hs00175798_m1 TRPM3 Hs00257553_m1 TRPM8 Hs01066596_m1 TRPV1 Hs00218912_m1 SCN9A (Nav1.7) Hs01076699_m1 SCN10A (Nav1.8) Hs01045137_m1 GAPDH Hs99999905_m1 OPRM1 (μ‐opioid receptor) Hs01053957_m1 HPRT1 Hs01003267_m1 Isl‐1 Hs00158126_m1 NTRK1 (TrkA) Hs01021011_m1 NTRK2 (TrkB) Hs00178811_m1 PRDM12 Hs00964106_m1 NANOG Hs02387400_g1 CALCA (CGRP) Hs01100741_m1 POU5F1B Hs04995079_g1 RUNX1 Hs01021970_m1 TAC1 Hs00243225_m1 TRPV2 Hs00901640_m1 TRPV3 Hs00376854_m1 TRPV4 Hs01099348_m1 TRPV5 Hs00219765_m1 TRPV6 Hs00367960_m1 TRPM1 Hs00170127_m1 TRPM2 Hs01066071_m1 TRPM4 Hs00214167_m1 TRPM5 Hs00175822_m1 TRPM6 Hs00214306_m1 TRPM7 Hs00292383_m1 TRPC1 Hs00608195_m1 TRPC2 Hs03453918_m1 TRPC3 Hs00162985_m1 TRPC4 Hs01077392_m1 TRPC5 Hs00202960_m1 TRPC6 Hs00175753_m1 TRPC7 Hs00220638_m1 TRPML1 Hs01100653_m1 TRPML2 Hs00401920_m1 TRPML3 Hs00217663_m1 TRPP1 Hs00165517_m1 TRPP2 Hs00175850_m1 TRPP3 Hs00950467_m1 Open in a separate window TaqManTM gene expression assays used in this study 2.4.

Techniques: Gene Expression

Journal: Frontiers in Pharmacology

Article Title: Hypo-Osmotic Loading Induces Expression of IL-6 in Nucleus Pulposus Cells of the Intervertebral Disc Independent of TRPV4 and TRPM7

doi: 10.3389/fphar.2020.00952

Figure Lengend Snippet: TaqMan primers list.

Article Snippet: TRPC4 , Transient Receptor Potential Cation Channel, Subfamily C, Member 4 , Bt03214662_m1.

Techniques: Activation Assay

Journal: Frontiers in Pharmacology

Article Title: Hypo-Osmotic Loading Induces Expression of IL-6 in Nucleus Pulposus Cells of the Intervertebral Disc Independent of TRPV4 and TRPM7

doi: 10.3389/fphar.2020.00952

Figure Lengend Snippet: Primary screening: number of donors (min. n = 0; max. n = 3), in which the mRNA expression of all available TRP channel was detectable using TaqMan gene array.

Article Snippet: TRPC4 , Transient Receptor Potential Cation Channel, Subfamily C, Member 4 , Bt03214662_m1.

Techniques: Expressing

Journal: Frontiers in Pharmacology

Article Title: Hypo-Osmotic Loading Induces Expression of IL-6 in Nucleus Pulposus Cells of the Intervertebral Disc Independent of TRPV4 and TRPM7

doi: 10.3389/fphar.2020.00952

Figure Lengend Snippet: Secondary screening: number of donors (min. n = 0; max. n = 5), in which the mRNA expression of all selected TRP channels was detectable using TaqMan gene array and qPCR.

Article Snippet: TRPC4 , Transient Receptor Potential Cation Channel, Subfamily C, Member 4 , Bt03214662_m1.

Techniques: Expressing

Journal: Science Advances

Article Title: Cryo-EM structure of TRPC5 at 2.8-Å resolution reveals unique and conserved structural elements essential for channel function

doi: 10.1126/sciadv.aaw7935

Figure Lengend Snippet: ( A ) Side view of TRPC5’s pore region with chains A and C compared with TRPC4 (gray). The ion conduction pathway is shown as dots and mapped using HOLE with key amino acid residues labeled. ( B ) Pore radius along the central axis. The side chains of glycine form a narrow constriction at the selectivity filter. “INQ” motif forms the lower gate. ( C ) Side view of TRPC5 (blue) monomer subunit compared with TRPC4 (gray). Differences in the organizations of a linker (red arrowheads) of S5 and the small loop above the pore helix of the extracellular pore domain are enlarged. ( D ) Sequence of the mouse TRPC5 aligned to TRPC4 and other TRPC subfamily members (Clustal Omega) between S5 and S6 including the linker, pore helix, and pore loop. Regions corresponding to different extracellular pore domains are indicated by the red arrowheads. The two cysteines forming disulfide bonds, a conserved “LFW” motif, and the selectivity filter are highlighted. ( E to G ) Patch clamp recordings of TRPC5 mutants in response to channel activators Gd 3+ and EA.

Article Snippet: Primary antibodies against TRPC5 and TRPC4 were purchased from Sangon Biotech (Shanghai, China) and Proteintech (Wuhan, China), respectively.

Techniques: Labeling, Sequencing, Patch Clamp

Journal: Science Advances

Article Title: Cryo-EM structure of TRPC5 at 2.8-Å resolution reveals unique and conserved structural elements essential for channel function

doi: 10.1126/sciadv.aaw7935

Figure Lengend Snippet: ( A ) Superimposed side views of mouse TRPC5 subunit (blue) compared with other TRPC family members, including mouse TRPC4 [Protein Data Bank (PDB) ID code 5Z96, gray] , human TRPC3 (PDB ID 5ZBG, pink) , and human TRPC6 (PDB ID 5YX9, yellow) . ( B ) The conserved LFW motif in the pore helix from a π-π interaction that stabilizes the pore region. ( C ) Key pore loop disulfide bond between Cys 553 and Cys 548 in TRPC5 (black arrowhead) and the corresponding pore loop disulfide bond in TRPC4 that close to a linker (red arrowheads) of S5 and loop above the pore helix. This disulfide bond is not present in TRPC3 or TRPC6. ( D ) Differences in the organization of the S3 helix between the TRPC4/5 and TRPC3/6. The S3 helices of TRPC3 and TRPC6 are longer than those of TRPC5 and TRPC4. ( E ) Patch clamp recordings of wild-type (WT) TRPC5 and cysteine mutants in response to the reducing agent [dithiothreitol (DTT)] and channel activator (EA). ( F ) Localization of enhanced yellow fluorescent protein–tagged WT TRPC5 and cysteine mutants in HEK293 cells. ( G ) Quantification of cell surface expression of TRPC5 cysteine mutants ( n = 3). Mean values are shown as gray bars. By analysis of variance (ANOVA), there is no statistically significant difference. a.u., arbitrary units.

Article Snippet: Primary antibodies against TRPC5 and TRPC4 were purchased from Sangon Biotech (Shanghai, China) and Proteintech (Wuhan, China), respectively.

Techniques: Patch Clamp, Expressing

Journal: Science Advances

Article Title: Cryo-EM structure of TRPC5 at 2.8-Å resolution reveals unique and conserved structural elements essential for channel function

doi: 10.1126/sciadv.aaw7935

Figure Lengend Snippet: ( A to F and I ) The time course of whole-cell currents measured at +80 and −80 mV and representative I-V relationships in different conditions as labeled. TRPC5-C4motif, TRPC5 mutant carrying the TRPC4 motif ETKGLS; TRPC4-C5motif, TRPC4 mutant with the TRPC5 motif TRAIDEPNN; TRPC5-C4motif-C553A-C558A, TRPC5 mutant bearing the TRPC4 motif and double-cysteine mutations. ( G and H ) Time constants (τ) of channel activation and inactivation by 100 nM EA. τ values from each recording are plotted as dots. “∞” indicates that the current did not decay in the recording time frame. Mean τ values of each mutant are shown as gray bars. There is no significant difference by ANOVA in (G) ( n = 8 to 11).

Article Snippet: Primary antibodies against TRPC5 and TRPC4 were purchased from Sangon Biotech (Shanghai, China) and Proteintech (Wuhan, China), respectively.

Techniques: Labeling, Mutagenesis, Activation Assay

Journal: Science Advances

Article Title: Cryo-EM structure of TRPC5 at 2.8-Å resolution reveals unique and conserved structural elements essential for channel function

doi: 10.1126/sciadv.aaw7935

Figure Lengend Snippet: ( A ) A cation (purple sphere) on the cytosolic face is in the hydrophilic pocket of the S1-S4 domain, interacting with Glu 418 , Glu 421 , Asn 436 , and Asp 439 . Enlarged view of the cation binding site. ( B ) Comparable Ca 2+ (green sphere) binding site and an enlarged view of TRPM4 as “E/Q/N/D” (TRPC5, “E/E/N/D”). ( C ) Sequence of the mouse TRPC5 aligned to mTRPC4 and other representative TRP members (Clustal Omega). The key residues are highlighted as E/E/N/D, which are conserved in TRPC2/3/5/6/7 and TRPM7 but as E/Q/N/D in TRPC4 and TRPM4. ( D ) Patch clamp recordings of TRPC5 mutants in response to the channel activators Gd 3+ and EA. For the N436R mutant, 12 transfected cells showed no response to EA; weak activation was observed in three cells.

Article Snippet: Primary antibodies against TRPC5 and TRPC4 were purchased from Sangon Biotech (Shanghai, China) and Proteintech (Wuhan, China), respectively.

Techniques: Binding Assay, Sequencing, Patch Clamp, Mutagenesis, Transfection, Activation Assay