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Table 1 Summary of treatments for evaluating HSP70 concentration, nitric oxide production and cell proliferation

From: Effect of in-vitro heat stress challenge on the function of blood mononuclear cells from dairy cattle ranked as high, average and low immune responders

Heat Treatment Assay Type
HSP70 Production NO Production Cell Proliferation
TN (37 °C) Plate remains in 37 °C until immediately after HS1 (22 h 37 °C) (collect @ same time as HS1 plate) Plate remain at 37 °C until 48 h after LPS stimulation Plate remain at 37 °C until 72 h after ConA stimulation
HS1(1 × 42 °C for 4 h) All plates, but TN, removed from 37 °C after overnight incubation and placed at 42 °C. Samples collected immediately after 4 h 18 h 37 °C + 4 h 42 °C). Other plates returned to 37 °C All plates, but TN removed from 37 °C after overnight incubation and placed at 42 °C for 4 h, then returned to 37 °C All plates, but TN removed from 37 °C after overnight incubation and placed at 42 °C for 4 h, then returned to 37 °C
18-h HS1 Plate removed from 37 °C 18 h after HS1 treatment and samples collected (18 h 37 °C + 4 h 42 °C + 18 h 37 °C) Not applicable Not applicable
HS2(2 × 42 °C for 4 h) Remaining 2 plates removed from 37 °C and placed at 42 °C. Samples collected from HS2 plate immediately after 4 h (18 h 37 °C + 4 h 42 °C + 18 h 37 °C + 4 h 42 °C). 18-h HS2 plate returned to 37 °C Only HS2 plates removed from 37 °C, following another overnight incubation, and placed at 42 °C for 4 h. Then return to 37 °C until 48 h after LPS stimulation when samples from all plates collected Only HS2 plates removed from 37 °C, following another overnight incubation, and placed at 42 °C for 4 h. Then return to 37 °C until 72 h after Con A stimulation when MTT assay for all plates performed.
18-h HS2 Plate removed from 37 °C 18 h after HS2 treatment and samples collected (18 h 37 °C + 4 h 42 °C + 18 h 37 °C + 4 h 42 °C + 18 h 37 °C) Not applicable Not applicable