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