Table 1 Relative NLRP3 mRNA expression in baseline and stimulated THP-1 and ARPE-19 cells.

	Vehicle	LPS	LPS + ATP	LPS + IL1α	LPS + IL1α + 7KCh	TNFα	Pam3CSK4	ox-LDL	LPS + TCDD	IL-17	THP-1 Veh	THP-1 (LPS + ATP)
Mean fold change	1.0	0.9	1.0	1.9	0.8	0.9	0.5	0.4	0.4	0.5	719	1019
SEM	0.0	0.3	0.4	1.0	0.7	0.6	0.3	0.3	0.2	0.2	25.3	62
Avg fold change exp #1	1.0	0.7	1.7	0.8	0.0	0.2	0.1	0.2	0.0	0.7
Avg fold change exp #2	1.0	0.5	0.4	1.0	0.3	0.3	0.5	0.1	0.4	0.1	744.8	1081.1
Avg fold change exp #3	1.0	1.5	0.8	3.9	2.1	2.1	0.9	1.0	0.6	0.7	694.3	957.2
	Ct Values
Experiment 1 NLRP3	35.9	36.9	36.0	41.0	39.8	41.0	41.0	37.1	41.0	36.8
	41.0	35.2	36.9	36.2	38.2	41.0	36.7	41.0	41.0	37.1
	37.0	37.2	34.8	35.8	39.8	35.8	41.0	41.0	38.1	36.7
Exp.1 reference RNA	26.4	24.7	25.8	26.0	23.0	25.3	24.1	26.0		24.6
	26.3	24.2	25.0	25.8	22.4	25.0	23.9	25.6	23.4	24.6
		23.7	24.1	25.7	22.0	25.0	23.7	25.3	23.7	24.8
Experiment 2 NLRP3	36.5	37.1	37.3	41.0	38.6	41.0	37.1	41.0	41.0	41.0	30.9	30.6
	36.9	41.0	37.0	37.0	39.4	36.8	41.0	41.0	41.0	41.0	31.2	31.1
	37.1	36.7	41.0	36.3	38.7	41.0	37.1	41.0	41.0	41.0	30.2	30.2
Exp.2 reference RNA	18.3	18.7	18.6	19.5	18.6	19.4	18.9	19.2	21.0	18.8	21.4	21.9
	18.2	18.6	18.6	19.5	19.0	19.4	19.0	19.1	21.0	18.9	22.0	22.3
	18.2	18.6	18.6	19.5	18.7	19.4		19.1	21.1	18.8	21.4	21.7
Experiment 3 NLRP3	39.0	41.0	39.8	37.9	37.5	37.6	39.9	38.9	41.0	39.9	28.5	28.2
	41.0	37.3	41.0	38.6	41.0	38.9	41.0	41.0	41.0	41.0	28.4	28.1
	39.1	38.1	41.0	38.8	37.3	39.0	41.0	39.1	38.9	39.9	28.3	27.9
Exp.3 reference RNA	20.9	20.6	21.9	21.6	20.9	20.9	21.8	21.0	21.0	21.9	19.3	19.2
	21.0	21.0	21.1	21.8	21.0	21.0	21.6	20.8	20.8	20.5	19.0	19.2
	20.9	20.3	21.7	21.4	20.9	20.5	21.9	20.9	20.8	20.8	19.3	19.3

Expression levels of NLRP3 mRNA measured in baseline and stimulated THP-1 and ARPE-19 cells were analyzed by RT-PCR. THP-1 cells were primed with LPS (10 μg/mL) plus ATP (5 mM) for 4 hours. ARPE-19 cells were stimulated with NLRP3-priming stimuli including LPS (10 μg/mL) alone, LPS (10 μg/mL) plus ATP (5 mM), LPS (10 μg/mL) plus TCDD (10 nM), LPS (20 μg/mL) plus IL-1α (25 ng/mL), LPS (20 μg/mL) plus IL-1α (25 ng/mL) plus 7-Ketocholesterol (10 μM), TNFα (10 ng/mL), IL-17α (100 ng/mL), Pam3CSK4 (300 ng/mL) for 24 hours and oxidized-LDL (500 μg/mL) for 48 hours. In ARPE-19 samples, expression levels are shown as fold changes in mRNA, relative to ARPE-19 vehicle from three biologically independent experiments (n = 3), that were performed in triplicates. Values expressed as mean fold changes in expression; 2^−ΔΔCt (2⁻(ΔCt - ΔCtvehicle)) ± standard error of the mean (SEM) and threshold cycle (Ct) values.

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