Fig. 5

RARRES2 negatively regulates the PTEN-mTOR-SREBP1 axis. a Western blotting analysis of the levels of PTEN, Akt, p-Akt (Ser473), mTOR, p-mTOR (Ser2448), SREBP1 and Chemerin in RARRES2-OE, shRARRES2 and control MDA-MB-231 cells. b Viability of shRARRES2-2 and control MDA-MB-231 cells treated with 0.25 μmol/L rapamycin. Two-way ANOVA with Geisser-Greenhouse correction: ^*P = 0.0017. c Rapamycin treatment (0.2 μmol/L) rescues cell invasion in shRARRES2-2 MDA-MB-231 cells (Scale bar = 100 μm). Two-way ANOVA with Sidak’s multiple comparisons test: ^**P = 0.0133. d Western blotting analysis of the levels of mTOR, p-mTOR (Ser2448) and SREBP1 in shRARRES2 and control MDA-MB-231 cells treated with 0.2 μmol/L rapamycin. e Western blotting of total lysates of RARRES2-OE and control MDA-MB-231 cells, which were transfected with short interfering RNA against CMKLR1 (si1 – 3) or a negative-control RNA (siNC). f mRNA levels of CMKLR1 and FASN in the cultures described in panel (e). Levels of mRNAs were normalized to those of GAPDH. ^***P < 0.001 (one-way ANOVA followed by Dunnett’s multiple-comparisons test). RARRES2 retinoic acid receptor responder 2, mTOR mammalian target of rapamycin, SREBP sterol regulatory element-binding protein, CMKLR1 chemokine-like receptor-1, FASN fatty acid synthase, RARRES2-OE RARRES2 overexpression, ns non-significant