Fig 2. Conditions of freezing and thawing for LoFT cell extraction.
(A) Effects of freezing temperature on protein concentration (protein conc.) after freeze-thawing. Samples were frozen in a deep freezer (-80°C) for 1 h, immersed in liquid nitrogen (Liq. N2) for 15 min, or not frozen. (B) Protein concentration (gray bars) in the LoFT cell extracts and volume of supernatant (Sup.) after centrifugation (circles) compared to the number of freeze-thaw cycles. Collectable supernatant volumes were derived from the viscosity of cell debris after freeze-thaw. Because the standard deviations of the collectable solution (n = 3) were smaller (on the scale shown) than the circle sizes, error bars are not indicated. All samples in (A) and (B) were thawed by incubation in ice water, and error bars indicate standard deviation (n = 3).