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An anti-β-actin antibody (sc47778) was purchased from Santa Cruz. Immunofluorescence staining Equal amounts of PASMCs were seeded in chamber well slides and then exposed to hypoxia or transfected with control mimic, miR-92b-3p or anti-miR-92b-3p. Cells were fixed in 2% paraformaldehyde, ...
The expression of hTSC1 gene was subsequently confirmed by western blot analysis using TSC1 specific antibody. As shown in Fig. 1c, the expression of TSC1 protein increased approximately 30% in multiple tissues from Tsc1 tg mice, including the heart, kidney, liver, and skeletal muscle. ...
HeLa cells with siRNA-mediated knockdown of TSC2 compared to a control (siC) were serum starved for 16 hr and immunofluorescently labeled with a TSC2 antibody. An enlarged view of a single cell is shown below. (E) Knockdowns of TBC1D7 and TSC1 do not have gross effects on the ...
The specimens were incubated with primary antibodies for 1 h at room temperature, followed by incubation with biotinylated anti-rabbit secondary antibody and streptavidin-horseradish peroxidase. 3, 3'-Diaminobenzidine was used as a chromogen and hematoxylin was used for counterstaining. The Phospho-4E...
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Antibodies to ATF4, CHOP, XBP-1, actin, U1 SnRNP 50, GAPDH, raptor and TSC2 were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Anti-ATF6 antibody was purchased from IMGENEX (San Diego, CA, USA), and anti-Rheb antibody was from ProSci (Poway, CA, USA). CBP epitope...
The following antibodies were used: TSC2 and actin (Santa Cruz Biotechnology); TSC1, MYC, p-PAK2, p-p70S6K and p-S6 (Cell Signaling); Rheb antibody was kindly provided by Richard Lamb (Department of Oncology, University of Alberta, Cross Cancer Institute, Edmonton, Alberta, Canada). ...
The mTORC1 complex was isolated and purified from HCT116 cells pretreated with MLN4924 (1 mM), rapamycin (100 nM, as positive control) or DMSO vehicle control for 24 h by immunoprecipitation using anti-mTOR antibody. A kinase reaction was initiated by incubating bead-conjugated mTORC1 complex ...
Full-length TSC2 and the M1028V5 and M1453V5 truncation proteins were immunoprecipitated with a polyclonal antibody recognising epitopes encoded by the last exon of TSC2 [5] and the washed immunoprecipitates were analysed by immunoblotting. In each case, the A1153V5 truncation protein was co...