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. 2020 Apr 12;9(4):951. doi: 10.3390/cells9040951

Figure 2.

Figure 2

Effect of MβCD-induced PM modulation on development of acquired thermotolerance (ATT) in B16-F10 cells. (A) B16-F10 cells were exposed for 10 min to 10 mM MβCD at 37 °C followed by 30, 60, or 90 min of heat stress at 42 °C (pre-exposure). Then, after 16–18 h recovery at 37 °C, cells were re-exposed to heat stress for 30 min at 45 °C. The following day, the fraction of surviving cells was estimated with resazurin (n = 4, * p < 0.05); (B) B16-F10 cells were incubated for 10 min with 10 mM MβCD at 37 °C followed by 60 or 90 min heat shock at 42 °C in serum-supplemented- or serum-free RPMI medium (limiting cholesterol supply). After pre-exposure heat stress, serum-free medium was exchanged for complete medium and cells were allowed to recover for 16–18 h at 37 °C. Then, cells were exposed for 30 min at 45 °C and the following day, the fraction of surviving cells was estimated with resazurin (n = 3, # p = 0.06, * p < 0.05); (C) B16-F10 cells were exposed for 10 min to 10 mM MβCD at 37 °C followed by 30, 60, or 90 min heat shock at 42 °C and cholesterol levels were measured immediately after heat stress (n = 6, * p < 0.05 compared to 37 °C control,). A.U.: arbitrary units.