Fig 2: Kidney-derived c-kit+ progenitor/stem cells modulate mTOR pathway in a time- dependent manner and up regulate Rictor in podocytes. (A) qPCR analyses of mTOR pathway in podocytes calculated by normalizing the data to WT-1 levels. Rictor and Raptor levels were down regulated in all groups independently of the time when compared to the normal kidney, except mTOR expression in the c-kit treated group at day 21. Rictor expression was higher in the c-kit treated group in comparison to the saline group at day 21. (B) Representative Western blots showing the levels of WT-1 in kidney cortex according to the treatment and time in the saline, c-kit and MSC-treated groups and in normal kidney. (C) Representative Western blots of mTOR pathway according to the treatment and time in the saline, c-kit and MSC-treated groups. (D) mTOR, Rictor, Raptor, phospo mTOR 2448 and 2481, and GβL increased in the c-kit group from day 10 to day 21 (all normalized to WT-1). When compared to the saline group at day 21, c-kit treated group exhibited higher levels of Rictor and phospho mTOR 2481, and Raptor as well, but not phospho mTOR 2448. Comparison was done accordingly to the normal kidney (set as 1), measured as arbitrary units (A.U.). *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001. Error bars represent means ± SEM. N = 3 in saline group and n = 4 in c-kit group at day 10 and n = 3 in saline, c-kit and MSC-treated groups at day 21.

Fig 3: Adipocytic GS overexpression via altered histone methylation induces chemoresistance to 5FU in peritoneal carcinomatosis of colorectal cancer metastasis. Adipocyte-derived glutamine (Gln) promotes resistance to 5FU chemotherapy via inducing mTOR activation in CRC cells. Adipocyte-derived Gln production is increased by Gln synthetase (GS) upregulation, which is regulated by altered histone methylation. Loss of H3k4me2 increases GS expression in adipose cells.

Fig 4: Tumor cells outcompete adipose cells for glutamine via mTORC1. (A) The protein levels of mTOR and p-mTOR (ser2448) in AD, B16, CT26, and MC38 cells were measured with Western blot. (B) The relative protein levels of mTOR and p-mTOR in (B) were calculated (n = 3, *P < 0.05, **P < 0.01). (C) Cell viability of CT26 cells treated with CM CM+5FU (10 μmol/L), CM+Rapamycin, or CM+5FU+Rapamycin by CCK8 assays (n = 5). (D) Mice were intraperitoneally inoculated with CT26 cells which were treated with 5FU, Rapamycin, or 5FU+Rapamycin, respectively. Then, the representative images of the mice-bearing tumors were shown. Arrow heads, tumors. (E) The tumors in (D) were isolated and weighed (n = 5, **P < 0.01). (F) Kaplan-Meier curves for the survival of 5FU-treated or 5FU+Rapamycin -treated TGGS mice after MC38 tumor cell inoculation plotted against time (days after injection) (**P < 0.01).

Fig 5: Analysis of proteins related to Cdk5, Akt/GSK, and mTOR pathways. A Ratio of phospho/total protein levels for the two targets of Cdk5 MAP2 and MAP1b as quantitated from the MS/MS spectra. B–D Western blot analysis and quantification of proteins belonging to mTOR pathway, including the two phosphorylated forms of mTOR, p-mTOR2481 (auto-phosphorylated residue), and p-mTOR2448 (phosphorylated by Akt), in neurons wild-type and knockouts for DJ-1 protein (a representative gel and global densitometry analysis are shown). E Levels of proteins related to Akt/GSK pathway identified by mass spectrometry. F, G Western blot analysis and quantification of Akt and phospho-Akt in neurons dj1+/+ and dj1−/−. Western blot data are mean ± SD (N ≥ 3; t test; *p < 0.05). Full-length blots are presented in Figs. S3 and S4