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Fig. 4 | Stem Cell Research & Therapy

Fig. 4

From: Inhibition of mTORC1 signaling protects kidney from irradiation-induced toxicity via accelerating recovery of renal stem-like cells

Fig. 4

Rapamycin treatment reduced increase of pS6 expression in irradiated kidney tissues. a Increased expression of pS6 and pmTOR by irradiation reduced via rapamycin treatment. Protein lysates prepared from kidney tissues with (Rap) or without (Veh) rapamycin treatment at day 7 (d7) after irradiation. Expression of pS6, S6, pmTOR and mTOR (left panel) detected by western blotting. GAPDH used as a housekeeping control. Nonirradiated kidney tissues (CTL) used as controls. Expression of pS6 and S6 quantitated by ImageJ software and ratios of pS6/S6 and pmTOR/mTOR presented (right panel). b pS6 immunostaining in kidney tissues. Kidney tissues with or without rapamycin treatment harvested at day 7 post irradiation and fixed in 4% paraformaldehyde after irradiation and pS6 immunostaining performed as indicated in Methods (left panel). Bar = 50 μm. Imaging software used to analyze integrated optical density (IOD) of pS6 (right panel). Arrows represent pS6-positive staining. c Expression of LC3-I and LC3-II in kidney tissues. Protein lysates were prepared from kidney tissues at day 7 after irradiation. Expression of LC3-I and LC3-II (left panel) detected by western blotting. GAPDH used as housekeeping control. Vehicle treated-kidney tissues (Veh) used as controls. Expression of LC3-I and LC3-II quantitated by ImageJ software and ratio of LC3-II and LC3-I expression presented (right panel). *p < 0.05 vs Veh; ***p < 0.001 vs Veh. CTL nonirradiated renal tissues, GAPDH glyceraldehyde 3-phosphate dehydrogenase, IHC immunohistochemistry, pmTOR pshosphorylated mammalian target of rapamycin, TBI total body irradiation

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