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. 2019 May 24;11:4781–4796. doi: 10.2147/CMAR.S207257

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© 2019 Fei et al.

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Eupatilin disrupts the cytoskeleton and affects intracellular relocalization of cofilin via the p-LIMK/cofilin pathway. (A) Treatment of U251MG cells with 160 μM eupatilin for 24 hrs and photographing with an inverted microscope. (B) F-actin was stained with Phalloidin-iFluor 488, and the microfilament shape of each layer (0.4 um) and superimposed layer (2.0 um) was observed by confocal microscopy, DAPI: 405 nm. (C) Immunofluorescence detection of Golgi marker protein GM130 (wavelength: 647 nm). (D) The expression of G-actin was detected by immunofluorescence (wavelength: 555 nm), and the optical density value (E) was calculated and statistically analyzed with image J. (F) Western detection of protein expression in the p-LIMK/cofilin pathway after treatment of U251MG with different time gradients and eupatilin concentration gradients. GAPDH is used as a housekeeping protein to prove the equal loading in each lane of the electrophoresis and to normalize densitometric values of the other protein analyzed. (G) Statistical analysis of Cofilin, p-cofilinSer3, and cofilin/p-cofilinSer with Quantity One after treatment with 160 μM eupatilin for different times, and the difference was statistically significant. *P<0.05 vs the control group. U251MG was treated with 160 μM eupatilin for 24 hrs, and p-cofilinSer3 (I) and cofilin (H) were stained to observe intracellular localization (wavelength: 647 nm). Each experiment was repeated three times.