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. 2021 Nov 17;13(22):5757. doi: 10.3390/cancers13225757

Figure 6.

Figure 6

Figure 6

PCAIs activate the MAPK pathway in A549 lung cancer cells. A549 cells were treated for 48 h with 0−5 μM of NSL-YHJ-2-27 or 10 μM of NSL-YHJ-2-62. These were then lysed and subjected to western blot analysis for total and phosphorylated levels of MAPK pathway proteins. (A) Shows treated cells captured using the Nikon Eclipse Ti 100 inverted microscope at 10× magnification just before lysis for western blotting analysis. (B-F) Western blot images and densitometry plots of bands following quantification using Image Lab Software normalized against GAPDH or α-Actinin. The samples were analyzed for total (B) BRaf (t-BRaf) and phosphorylated BRaf (p-BRaf); (C) total CRaf (t-CRaf) and phosphorylated CRaf (p-CRaf); (D) total MEK1/2 (t-MEK1/2) and phosphorylated MEK1/2 (p-MEK1/2); (E) total ERK1/2 (t-ERK1/2) and phosphorylated ERK1/2 (p-ERK1/2); and (F) total p90RSK (t-p90RSK) and phosphorylated p90RSK (p-p90RSK). Data are representative of three independent experiments. Statistical significance (* p < 0.05 and ** p < 0.01) was determined by 1-way ANOVA with post hoc Dunnett’s test. Original blots see Figure S1.