Figure 5.

Effect of the specific p38 MAPK activator, anisomycin, (see “Materials and Methods”) and the effect of stearic acid (SA) on (A) the level of p38 MAPK, phospho-p38 MAPK and phospho-MAPKAPK-2 (substrate of p38 MAPK); (B) the level of phospho-c-Raf, phospho-MEK1/2, phospho-ERK1/2 (the ERK signaling pathway); (C) cell growth and viability; and (D) cleavage of PARP, caspase-7 (C7), caspase-8 (C8) and caspase-9 (C9) (markers of apoptosis) in NES2Y cells. Cells incubated without the activator and SA represented control cells. After 12 h of incubation (see “Materials and Methods”) (A,B,D), the level of individual proteins was determined using western blot analysis and the relevant antibodies (see “Materials and Methods”). A monoclonal antibody against human actin was used to confirm equal protein loading. The data shown were obtained in one representative experiment from three independent experiments. When assessing cell growth and viability (C), 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 ± SEM. * p < 0.05 when comparing the number of control cells and cells with anisomycin or SA.