Journal: Scientific Reports

Article Title: Rottlerin-mediated inhibition of Toxoplasma gondii growth in BeWo trophoblast-like cells

doi: 10.1038/s41598-017-01525-6

Figure Lengend Snippet: T. gondii and Rottlerin induce morphological and biochemical alterations associated with autophagy in BeWo Cells. Left panel: control-vehicle treated cultures ( A , B and C ); 5 µM Rottlerin-treated cultures ( D , E and F ); T. gondii infected cultures ( G , H and I ); T. gondii infected and 5 µM Rottlerin treated cultured ( L , M , N and O ). Arrows: cytoplasmic vacuoles; star: T. gondii ; double arrow head: expansion of the rough endoplasmic reticulum; arrow head: double membraned vacuoles ( P and Q ): representative western blot and densitometric analysis performed on three independent experiments of LC3I/II ( P ) and p62 ( Q ) proteins in BeWo cells after infection and treatment with 5 µM Rottlerin. *p < 0.05; # p < 0.01. ( R ): representative western blot of three independent experiments of PARP proteins in BeWo cells after infection and treatment with 5 µM Rottlerin. ( S ): viability of BeWo cells after infection and treatment with 5 µM Rottlerin (Rt). ( A ) Bar = 5 µm; ( B , C , F and I ), Bar = 500 nm; ( D , L and M ), Bar = 2 µm; ( E , G , H ), Bar = 1 µm.

Article Snippet: Then, the blots were probed with polyclonal antibodies anti human LC3I/II, p4EBP-1, Beclin 1, peIF2α and PARP (Cell Signaling Technology, Boston, MA USA), SQSTM1/p62 and β-actin (Santa Cruz Biotechnology; Santa Cruz, CA) at dilution of 1:1000 overnight at 4 °C.

Techniques: Infection, Cell Culture, Western Blot