Fig. 3

SGK1 inhibition induces autophagy, which contributes to metastasis suppression. a-e PC3 and LNCaP cells were infected with empty lentivirus control (LV2-Ctrl) or with the lentivirus SGK1-shRNA (LV2-shSGK1) and subsequently incubated for 24 h before pretreatment with 5 mM 3MA or PBS (control). Autophagy activity was determined with AO staining (a) and quantified (b) in LNCaP cells. c Wound healing assays of PC3 cells. The wound healing area was analyzed by using ImageJ software and the corresponding data, relative to 0 h, expressed in the graph (d). e Matrigel cell invasion assays were performed both in PC3 and LNCaP cells. Representative images show the cells that invaded through the matrigel. Representative histograph of invaded tumor cells is displayed and number of invaded tumor cells quantified. f PC3 and LNCaP cells were infected with empty lentivirus control (LV2-Ctrl) or with the lentivirus SGK1-shRNA (LV2-shSGK1) and subsequently incubated for 48 h before pretreatment with 5 mM 3MA or PBS (control). Western blots were probed for levels of SGK1, LC3-I, LC3-II, MMP3 and MMP9, and GAPDH was used as a loading control. All results are representative of three experiments and are expressed as the mean ± S.D.; *P < 0.05