Table 1. Changes in TRPV4 sparklet sites per field and average number of open TRPV4 channels (NP_O) per site.

CPA (30 µM) was included under all conditions. The PLC inhibitor U73122 (5 µM) was added 10 min before the addition of CCh. The PKC inhibitor Gö-6976 (1 µM) was added 10 min before CCh (10 µM), OAG (10 µM), or PMA (10 nM). The TRPV4 inhibitor HC-067047 (1 µM) was added after CCh, OAG, or PMA for 10 min. The data in the presence of GSK101 (3 nM) represent sparklet sites at MEPs. Significant differences were assessed by paired t test or one-way ANOVA with post hoc Bonferroni test. *P < 0.05; ns, not significant (versus basal or GSK101, unless specified).

Stimulation conditions	Active sites per field	NPO per site	n	Significant difference
Basal	2.2 ± 0.4	0.05 ± 0.02	10
+ CCh	5.0 ± 0.8	0.15 ± 0.02	10	*
+ CCh + HC-067047	2.0 ± 0.4	0.03 ± 0.01	5	* (versus CCh)
Basal + U73122	2.2 ± 0.2	0.04 ± 0.02	5
+ CCh	2.4 ± 0.3	0.06 ± 0.02	5	ns
Basal	3.1 ± 0.4	0.04 ± 0.01	8
+ OAG	4.8 ± 0.2	0.12 ± 0.02	8	*
+ OAG + HC-067047	1.6 ± 0.4	0.05 ± 0.01	5	* (versus OAG)
Basal	2.6 ± 0.4	0.05 ± 0.01	8
+ PMA	4.9 ± 0.8	0.13 ± 0.02	8	*
+ PMA + HC-067047	1.8 ± 0.4	0.04 ± 0.01	4	* (versus PMA)
Basal + Gö-6976	2.2 ± 0.4	0.06 ± 0.02	9
Gö-6976 + CCh	2.8 ± 0.3	0.04 ± 0.01	9	ns
Basal + Gö-6976	3.4 ± 0.6	0.06 ± 0.02	9
Gö-6976 + OAG	3.1 ± 0.6	0.05 ± 0.02	9	ns
Basal + Gö-6976	2.5 ± 0.2	0.05 ± 0.01	9
Gö-6976 + PMA	3.6 ± 0.6	0.03 ± 0.01	9	ns
GSK101, MEPs	6.4 ± 0.7	0.07 ± 0.02	7
+ CCh	14.8 ±1.4	0.16 ± 0.03	7	*
+ CCh + HC-067047	2.0 ± 0.6	0.08 ± 0.02	4	* (versus CCh)
GSK101, MEPs	7.1 ±1.0	0.09 ± 0.02	4
+ OAG	17.1 ±1.6	0.17 ± 0.03	4	*
+ OAG + HC-067047	4.0 ± 0.8	0.05 ± 0.01	3	* (versus OAG)
GSK101, MEPs	6.3 ±1.2	0.08 ± 0.01	5
+ PMA	12.0 ± 2.4	0.19 ± 0.02	5	*
+ PMA + HC-067047	3.3 ± 0.3	0.04 ± 0.01	3	* (versus PMA)
MEPs
GSK101 + Gö-6976	6.8 ±1.3	0.07 ± 0.02	7
+ CCh	6.9 ±1.0	0.07 ± 0.03	7	ns
GSK101 + Gö-6976	7.4 ±1.6	0.08 ± 0.03	4
+ OAG	8.1 ±1.9	0.08 ± 0.02	4	ns
GSK101 + Gö-6976	6.5 ±1.2	0.06 ± 0.02	3
+ PMA	7.6 ±1.6	0.07 ± 0.01	3	ns