Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: Short-Term Stimulation of Calcium-Permeable Transient Receptor Potential Canonical 5–Containing Channels by Oxidized Phospholipids

doi: 10.1161/atvbaha.110.205666

Figure Lengend Snippet: Figure 6. Role of Gi/o signaling but not PAF receptors. Data are from intracellular Ca2 measurements from TRPC5- expressing HEK 293 cells (A-C) and VSMCs (D-F). Cells were pretreated with 1-g/mL pertussis toxin (PTX) or dena- tured (denat) PTX for 4 hours at 37°C. A through C, Responses were evoked by 3-mol/L PGPC (A, n/N3/9), 5-mol/L POVPC (B, n/N3/9), or 5-mol/L lyso- phosphatidylcholine (C, n/N3/18). D, A representative paired experiment showing responses to 3-mol/L PGPC after treat- ment with PTX or denatured PTX (N5 each). E, Responses were evoked by 3-mol/L PGPC or 5-mol/L POVPC (n/N4/20 each). F, Responses to 3-mol/L PGPC in cells preincubated and maintained in the presence of 100-mol/L WEB2086 or its vehicle (N6 each and representative of 3 independent experiments).

Article Snippet: Stocks of prostaglandin-E2 (PGE2) and ( )-15-deoxy-16R-hydroxy-17-cyclobutyl PGE1 ((R)-butaprost) at 10 mmol/L and 3-(4-[2-chlorophenyl]-9-methyl-6Hthieno[3,2-f](1,2,4)triazolo[4,3- ]diazepine-2-yl)-1-propanone (WEB2086; Tocris Bioscience) at 100 mmol/L were prepared in ethanol.

Techniques: Expressing

Journal: Arteriosclerosis, thrombosis, and vascular biology

Article Title: Biphasic regulation of transcription factor nuclear factor-kappaB activity in human endothelial cells by lysophosphatidylcholine through protein kinase C-mediated pathway.

doi: 10.1161/01.atv.18.4.568

Figure Lengend Snippet: Figure 9. Effect of PAF-receptor blocker on lysoPC-induced endothelial NF-kB activation. HUVECs were incubated for 60 minutes with lysoPC (15 mmol/L) in the presence of WEB 2086 (5.0 or 10.0 mmol/L), which is a specific PAF-receptor blocker. After the incubation, the nuclear proteins were extracted and analyzed by EMSA (7.5% gel). Arrow indicates the bands of NF-kB/DNA complex. Lane 1 (control), treated with vehicle; lane 2, treated with lysoPC alone; lane 3, treated with WEB 2086 (5.0 mmol/L)1lysoPC; lane 4, treated WEB 2086 (10.0 mmol/ L)1lysoPC. The result shown is representative of four indepen- dent experiments.

Article Snippet: Antibodies raised against p50, p65, RelB, and c-Rel were from Santa Cruz Biotechnology Inc. PAF receptor blocker (WEB 2086) was a gift from Boehringer Mannheim (Mannheim, Germany).

Techniques: Activation Assay, Incubation, Control

Journal: iScience

Article Title: Macrophage/microglia-producing transient increase of platelet-activating factor is involved in neuropathic pain

doi: 10.1016/j.isci.2024.109466

Figure Lengend Snippet: Difference in the effect of intrathecal WEB2086 injection against neuropathic mechanical allodynia (A and B) Paw withdrawal threshold (PWT) to mechanical stimuli is assessed by the von Frey test and calculated by the up-down method. WEB2086 (a PAF receptor antagonist, 1–10 nmol) was administrated intrathecally (A) on day 7, (B) day 14 after nerve injury (n = 5–7). Arrows indicate the timing of administration. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001 vs. vehicle. Data are represented as mean ± SEM. See also Figure S2 .

Article Snippet: WEB2086 , Tocris Bioscience , Cat# 2339.

Techniques: Injection

Journal: iScience

Article Title: Macrophage/microglia-producing transient increase of platelet-activating factor is involved in neuropathic pain

doi: 10.1016/j.isci.2024.109466

Figure Lengend Snippet:

Article Snippet: WEB2086 , Tocris Bioscience , Cat# 2339.

Techniques: Recombinant, Software

Journal: Molecular and cellular endocrinology

Article Title: Role of cAMP/PKA pathway and T-type calcium channels in the mechanism of action of serotonin in human adrenocortical cells

doi: 10.1016/j.mce.2016.10.008

Figure Lengend Snippet: Coupling of 5-HT receptors to T-type Ca2+ currents in human cultured adrenocortical cells. (A) Superimposed Ca2+ currents elicited by increasing depolarizing pulses (from −60 mV to −30 mV) from a holding potential of −80 mV (upper panel). Current-voltage relationships of the peak Ca2+ currents (lower panel). (B) Superimposed Ca2+ currents elicited by depolarizing pulses from −80 mV to −50 mV and −40 mV recorded in the absence (control) or presence of 5-HT (10−5 M). (C) Effects of graded concentrations of 5-HT on cortisol secretion by adrenocortical cells cultured in the absence or presence of the T-type Ca2+ channel blockers mibefradil (10−6 M; ■) and NiCl2 (8 × 10−5 M; ●). (D) Effects of the L-type channel blocker nifedipine (10−5 M), N-type channel blocker ω-conotoxin (10−6 M) and P-type channel blocker sFTX-3.3 (10−6 M) on basal and 5-HT-induced cortisol production. BL, Basal level. *, p < 0.05; ***, p < 0.001 versus basal production in the absence of test substance. ###, p < 0.001 versus production in the presence of 5-HT.

Article Snippet: Serotonin, dibutyryladenosine 3′,5′-cyclic monophosphate (dbcAMP), 1-methyl-3-isobutylxanthine (IBMX), chelerythrine, H-89, GR113808, zacopride, EGTA, nifedipine, mibefradil, NiCl 2 were purchased from Sigma-Aldrich (Saint-Quentin Fallavier, France). ω-conotoxin and sFTX-3.3 were obtained from Alomone Labs (Jerusalem, Israel).

Techniques: Cell Culture