Fig 2: Raptor expression in ERα-positive, ERα-negative, and tamoxifen-resistant breast cancer cells. Raptor expression was higher in ERα-positive than in ERα-negative cell lines by Western blot analysis (a). Comparing ERα-positive with ERα-negative cells shows raptor in the ERα-positive but not in the ERα-negative nuclei as observed by in situ immunofluorescence (b). Raptor presence increased in the nucleus (N) in MCF7 tamoxifen-resistant cells (TamR) compared with MCF7 parental cells (Control), and changes in mTORC1 and Akt signaling were observed (c)

Fig 3: The mechanism underlying the RPTOR-promoted NSCLC-BM via the SPHK2/S1P/STAT3 signaling pathway. RPTOR regulated the transcriptional activation of the ceramidase SPHK2 by binding to the transcription factor YY1. SPHK2 catalyzed the degradation of ceramide to S1P. The latter, in turn, activated the STAT3 signaling pathway by binding to S1PR1 (solid lines), altering the microenvironment and enhancing the permeability of the BBB (dotted lines), resulting in the BM of NSCLC

Fig 4: The SASP is inhibited by rapamycin in endothelial cells (ECs) subjected to SIPS. a Real-time PCR analysis of SASP factors using cDNA derived from human coronary artery endothelial cells (HCAECs) at 72 h in Fig. 1a. The results are shown after normalization to values obtained from control HCAECs (value = 1). Results are presented as means ± SD from three independent experiments. ap < 0.05 vs. the Ctr and bp < 0.05 vs. the H2O2-treated HCAECs. b Immunoblotting for the levels of mTOR, RICTOR, RAPTOR, and MAPKAPK2 was performed using cell lysates and mTOR immunoprecipitates from HCAECs at 72 h in Fig. 1a. β-ACTIN was used as a loading control. The histogram shows mean densitometric readings for the proteins normalized to β-ACTIN or mTOR from three independent experiments. ap < 0.01 vs. the Ctr and bp < 0.01 vs. the H2O2-treated HCAECs. Control (Ctr): untreated cells. SIPS, stress-induced premature senescence. SASP, senescence-associated secretory phenotype

Fig 5: PAI reduces autophagic flux by promoting the mTOR complex pathways. (A) LM8 cells were loaded with Cyto-ID and treated with PAI (50%) or rapamycin (positive control) for 18 h and analyzed by flow cytometry. (B) LM8 cells were treated with PAI (50%) for the indicated time and then analyzed for the expression of p70-S6 kinase, Rictor, Raptor, p62, and LC3-I/II, and their phosphorylated forms by western blotting analysis using specific antibodies. GAPDH was used as a loading control. See Supplementary Figures 3 , 4 for examples of uncropped images and quantification for each antibody.