Table 2.

Determination of cytokine protein from C57BL/6J mouse brain homogenate spiked with known quantities of murine recombinant interleukin-1β (IL-1β), IL-6, or tumor necrosis factor α (TNFα). Three-plex assays were run to determine cytokine protein concentrations simultaneously. Cytokine protein detected in unspiked samples of whole brain homogenate represent basal cytokine expression in C57BL/6J mouse brain during this portion of the light:dark cycle. All samples were run in duplicate.

Protein in whole brain homogenate	Amount of recombinant cytokine added (IL-1β/IL-6/TNFα)	Recombinant IL-1β detected (% of spike)	Recombinant IL-6 detected (% of spike)	Recombinant TNFα detected (% of spike)
0 µg	0/0/0/ pg	ND	ND	ND
	5/5/5 pg	4 pg (80)	4 pg (80)	5 pg (100)
	20/20/20 pg	19 pg (95)	19 pg (95)	19 pg (95)
	50/50/50 pg	46 pg (92)	44 pg (88)	46 pg (92)
	500/500/500 pg	484 pg (97)	499 pg (100)	493 pg (99)
100 µg	0/0/0/ pg	14 pg	27 pg	22 pg
	5/5/5 pg	17 pg (89)	30 pg (94)	25 pg (93)
	20/20/20 pg	32 pg (94)	45 pg (96)	37 pg (88)
	50/50/50 pg	59 pg (92)	70 pg (91)	65 pg (90)
	500/500/500 pg	482 pg (94)	497 pg (94)	464 pg (93)
200 µg	0/0/0/ pg	26 pg	50 pg	41 pg
	5/5/5 pg	32 pg (103)	54 pg (98)	44 pg (96)
	20/20/20 pg	46 pg (100)	67 pg (96)	56 pg (92)
	50/50/50 pg	71 pg (93)	87 pg (87)	80 pg (88)
	500/500/500 pg	484 pg (92)	514 pg (93)	484 pg (89)

ND: not detected.