Figure 1.

Effect of 1 mM stearic acid (SA) (see “Materials and Methods”) on (A) cell growth and viability; (B) the level of cleaved PARP, caspase-7 (C7), caspase-8 (C8) and caspase-9 (C9) (markers of apoptosis); (C) the level of phospho-MKK3/6, p38 MAPK, phospho-p38 MAPK, phospho-MAPKAPK-2 (p38 MAPK signaling pathway); and (D) the level of phospho-c-Raf, phospho-MEK1/2, ERK1/2, phospho-ERK1/2 (the ERK signaling pathway) in NES2Y cells. Cells incubated without SA represented control cells. After 18 h of incubation (see “Materials and Methods”) for markers of apoptosis (B) and 3, 6, 12 and 24 h of incubation for p38 MAPK and ERK pathways members (C,D), the levels of individual proteins were determined using Western blot analysis and the relevant antibodies (see “Materials and Methods”). Monoclonal antibody against human actin was used to confirm equal protein loading. The data shown were obtained in one representative experiment from at least three independent experiments. When assessing cell growth and viability (A), cells were seeded at a concentration of 2 × 10⁴ cells/100 µL of culture medium per well of 96-well plate (see “Materials and Methods”). The number of living cells was determined after 48 h of incubation. Each column represents the mean of four separate cultures ± standard error of the mean (SEM). * p < 0.05 when comparing the number of control cells and cells treated with SA.