Figure legends
Figure 1 . Time courses (A and C) and concentration-effect
curves (B, D and E) in response to β-alanine in DMR (A, B and C) and
FLIPR assays (D) in CHO cells expressing recombinant human MRGPRD or
untransfected CHO cells (E).
Figure 2 . Time courses (A, B and C) and concentration-effect
curves (D) and (E) in response to β-alanine in DMR (A, B, C and D) and
FLIPR assays (E) in CHO cells expressing recombinant human MRGPRD. Cells
were preincubated with either YM-254890 (10µM) (A) PTX (0.1µg/ml) (B) or
PTX (0.1µg/ml) plus YM-254890 (10µM), (C) and concentration-response
curves determined (D). The effects of YM-254890, PTX and PTX plus
YM-254890 on FLIPR
Ca2+ assays were
also determined (D). Concentration-effect data are expressed as a
percentage of the maximum response to agonist and are the mean ± SEM of
a minimum of 5 replicates
Figure 3 . Time courses (A – D) for responses to GABA in the
DMR assay in the presence of vehicle only (A), in presence of the
Gq inhibitor YM254890 (10µM) (B), in the presence of PTX
(0.1µg/ml) (C), or in the presence of both the indicated concentrations
of YM-254890 and PTX (D).
Concentration-effect curves (E)
for these responses in the DMR assay at the 4-minute time point.
Concentration-effect curves (F) in the FLIPR Ca2+assay. Concentration-effect data are expressed as a percentage of the
maximum response to agonist and are the mean ± SEM of a minimum of 5
replicates.
Figure 4 . Time courses (A – D) for responses to Glycine in the
DMR assay in the presence of vehicle only (A), in presence of the
Gq inhibitor YM254890 (10µM) (B), in the presence of PTX
(0.1µg/ml) (C), or in the presence of both the indicated concentrations
of YM-254890 and PTX (D). Concentration-effect curves (E) for these
responses in the DMR assay at the 4-minute time point.
Concentration-effect curves (F) in
Ca2+ FLIPR
assay. Concentration-effect data are expressed as a percentage of the
maximum response to agonist and are the mean ± SEM of a minimum of 5
replicates.
Figure 5 . Time course and concentration-responses to the
Thioridazine hydrochloride response in the DMR assay in CHO cells
expressing the human MRGPRD receptor in the absence (A), and presence
(B) of PTX (0.1µg/ml) or in untransfected CHO cells (C).
Concentration-effect responses in FLIPR Ca2+ assay in
MRGPRD expressing CHO cells (D) and cell viability determined using the
CyQUANT assay over 72 hours (E).
Figure 6 . Concentration-effect curves in MRGPRD expressing CHO
cells in response to β-alanine (A, B), GABA (C, D) and Glycine (E, F) in
DMR (A, C, E) and FLIPR Ca2+ (B, D, F) assays, in the
presence of increasing concentrations of PD123,319. Data are expressed
as a percentage of the maximum response to agonist and are the mean ±
SEM of a minimum of 5 replicates
Figure 7 . Concentration-effect curves in MRGPRD expressing CHO
cells in response to β-alanine (A, B), GABA (C, D) and Glycine (E, F) in
DMR (A, C, E) and FLIPR
Ca2+ (B, D, F)
assays, in the presence of increasing concentrations of MU-6840. Data
are expressed as a percentage of the maximum response to agonist and are
the mean ± SEM of a minimum of 5 replicates
Figure 8 . Schild analyses (D, E, F) derived from
concentration-effect curves in response to β-alanine (A, B, C) in FLIPR
Ca2+ (A, B, D, E) and DMR (C, F) assays in the
presence of increasing concentrations of PD123,319 or MU-6840 (B, E).
Figure 9. Supplementary data. Time and concentration-effect in
response to increasing concentrations of PD123,319 (A) and MU-6840 (B)
in MRGPRD expressing CHO cells in the DMR assay. Concentration-effect
curves to increasing concentrations of Alamandine in FLIPR
Ca2+ assay in MRGPRD expressing CHO cells (C) and in
CyQUANT cell viability assay over 72 hours (D).