10.2337/db13-0970 [PubMed] [CrossRef] [Google Scholar] Biessels, G. mRNA manifestation levels of were analyzed by quantitative real\time PCR. = 5 from independent experiments with 2 technical replicates each. b The Bafetinib (INNO-406) cells were treated with D\glucose (25 mM) or Bafetinib (INNO-406) L\glucose (25 mM) for 24 h. Then, APP and \actin were detected by western blot. = 7 from independent experiments. * 0.05 vs. control, 0.05 vs. D\glucose. Quantitative data are presented as a mean S.E.M. All blots are representative. Figure S3. PICALM is located in lipid rafts under both normal\ and high glucose\conditions. a High glucose (25 mM) were treated for 24 h in the cells. Sucrose gradient\fractionized lysates were subjected to western blot. PICALM, CAV1, and FLOT1 were detected. Exploratory data. Figure S4. High glucose impairs autophagy in a time dependent manner. a The SK\N\MCs were treated with high glucose (25 mM) in a time dependent manner. LC3, P62, and \actin were subjected to western blot. = 5 from independent experiments. b The cells were treated with D\glucose (25 mM) or L\glucose (25 mM) for 24 h. Then, LC3, P62, and \actin were detected by western blot. = 5 from independent experiments. * 0.05 vs. control. Quantitative data are presented as a mean S.E.M. All blots are representative. Figure S5. PICALM induces autophagy under high glucose conditions. a, b The SK\N\MCs were treated with high glucose (25 mM) for 24 h. a Co\immunoprecipitation of PICALM with IgG and LC3 antibodies were shown in left panel. Total protein expressions in lysate were shown in right panel. = 5 from independent experiments. * 0.05 vs. Control. b Bafetinib (INNO-406) The cells were immunostained with VAMP2 or VAMP8 or VAMP3\specific antibodies and counterstained with DAPI. Scale bars, 8 m (magnification, 1,000). = 5 from independent experiments. Exploratory data. c The cells were transfected with NT siRNA Bafetinib (INNO-406) or siRNA for 12 h prior to high glucose (25 mM) treatment for 24 h. LC3 and \actin were detected by western blot. = 5 from independent experiments. Logarithmic transformations were performed for homogeneity of the sample variance. * 0.05 vs. NT siRNA transfection, # 0.05 vs. NT siRNA transfection + high glucose. Quantitative data are presented as a mean S.E.M. All blots and immunofluorescence images are FAZF representative. Figure S6. High glucose has no significant alterations Bafetinib (INNO-406) on the protein expression of Beclin1. The SK\N\MCs were treated with high glucose (25 mM) in a time dependent manner. The Beclin1 were detected by western blot. = 5 from independent experiments. Quantitative data are presented as a mean S.E.M. All blots are representative. Figure S7. High glucose has no significant alterations on the mRNA expression of endo\lysosomal fusion related proteins. a The SK\N\MCs were treated with high glucose (25 mM) in a time dependent manner. The mRNA expression levels of were analyzed by quantitative real\time PCR. = 5 from independent experiments with 2 technical replicates each. Quantitative data are presented as a mean S.E.M. Figure S8. The treatment of rapamycin attenuated STZ\induced phosphorylation of mTORC1. a Hippocampal sample was obtained from vehicle\, STZ\, STZ + rapamycin\, or rapamycin\treated mice. Then, p\mTOR (Ser 2,448), t\mTOR, and \actin were subjected to western blot. = 5 in each animal. * 0.05 vs. vehicle\treated mice, # 0.05 vs. STZ\treated mice. Quantitative data are presented as a mean S.E.M. All blots are representative. Table S1. Body weight and blood glucose levels in ZLC and ZDF rats. Blood glucose.