p mtor cat no 2971 antibodies  (Cell Signaling Technology Inc)

Journal: Experimental and Therapeutic Medicine

Article Title: Targeting translocator protein protects against myocardial ischemia/reperfusion injury by alleviating mitochondrial dysfunction

doi: 10.3892/etm.2024.12638

Figure Lengend Snippet: TSPO regulates autophagy and mitophagy via related pathways following A/R stimulation. (A) Representative fluorescence images and quantitative analysis of LC3B expression. Scale bar, 20 µm. (B) Western blotting and (C) densitometry analysis of LC3B, Beclin1 and ATG5 expression levels in si-NC and si-TSPO H9c2 cells with or without A/R stimulation. (D) Western blotting and (E) densitometry analysis of p-PI3K, PI3K, p-Akt, Akt, p-mTOR and mTOR expression levels. (F) Western blotting and (G) densitometry analysis of P62, PINK1 and Parkin expression levels. (H) Western blotting and (I) densitometry analysis of Mfn2 and Drp1 protein expression levels. * P<0.05, ** P<0.01, *** P<0.001, **** P<0.0001. TSPO, translocator protein; A/R, anoxia/reoxygenation; si, small interfering; NC, negative control; p-, phosporyalated.

Article Snippet: Membranes were blocked with 5% skimmed milk at room temperature for 1 h, then incubated with the following primary antibodies overnight at 4˚C: GAPDH (cat. no. HRP-60004; ProteinTech Group, Inc.), cleaved-caspase-3 (cat. no. 9661S; Cell Signaling Technology, Inc.), caspase-3 (cat. no. 66470-1-Ig; ProteinTech Group, Inc.), Bax (cat. no. 2772S; Cell Signaling Technology, Inc.), Bcl-2 (cat. no. 26593-1-AP; ProteinTech Group, Inc.), TSPO (cat. no. ab109497; Abcam), LC3B (cat. no. ab51520; Abcam), ATG5 (cat. no. 12994S; Cell Signaling Technology, Inc.), Beclin 1 (cat. no. 66665-1-Ig; ProteinTech Group, Inc.), P62 (23214S; ProteinTech Group, Inc.), PINK1 (cat. no. 23274-1-AP; ProteinTech Group, Inc.), Parkin (cat. no. 66674-1-Ig; ProteinTech Group, Inc.), Mfn2 (cat. no. 12186-1-AP; ProteinTech Group, Inc.), Drp1 (cat. no. 12957-1-AP; ProteinTech Group, Inc.), ATP5B (cat. no. 17247-1-AP; ProteinTech Group, Inc.), PI3K (cat. no. 4257S; Cell Signaling Technology, Inc.), phosphorylated (p)-PI3K (cat. no. 4228S; Cell Signaling Technology, Inc.), Akt (cat. no. 9272S, Cell Signaling Technology, Inc.), p-Akt (cat. no. 4060S; Cell Signaling Technology, Inc.), mTOR (cat. no. 2983S; Cell Signaling Technology, Inc.), p-mTOR (cat. no. 2971S; Cell Signaling Technology, Inc.), PKR-like ER kinase (PERK; cat. no. ab229912; Abcam), p-PERK (cat. no. 3179S; Cell Signaling Technology, Inc.), ATF6 (cat. no. 24169-1-AP; ProteinTech Group, Inc.) and inositol-requiring enzyme 1 (IRE1; cat. no. bs16696R, BIOSS).

Techniques: Fluorescence, Expressing, Western Blot, Negative Control

Journal: International Journal of Molecular Medicine

Article Title: TGIF1 plays a carcinogenic role in esophageal squamous cell carcinoma through the Wnt/β-catenin and Akt/mTOR signaling pathways

doi: 10.3892/ijmm.2021.4910

Figure Lengend Snippet: TGIF1 promotes the activation of the Wnt/β-catenin and Akt/mTOR signaling pathways. (A) Expression of Wnt/β-catenin-related and Akt/mTOR-related proteins were detected by western blot analysis in KYSE-150 and Eca-109 cells. (B) Quantitative analysis of the results of western blot analysis in KYSE-150 cells. (C) Quantitative analysis of the results of western blot analysis in Eca-109 cells. * P<0.05, vs. respective control. TGIF1, TGFB induced factor homeobox 1; mTOR, mammalian target of rapamycin.

Article Snippet: Anti-Bcl-2 (cat. no. 12789-1-AP), anti-Bax (cat. no. 50599-2-Ig), anti-cyclin 1 (cat. no. 60186-1-Ig) and anti-GAPDH (cat. no. 60004-1-lg) antibodies were obtained from Proteintech Group, Inc. Anti-cleaved caspase-3 (cat. no. 9661), anti-Wnt3a (cat. no. 2391), anti-β-catenin (cat. no. 9562), anti-Akt (cat. no. 9272), anti-p-Akt (cat. no. 9271), anti-mTOR (cat. no. 2972) and anti-p-mTOR (cat. no. 2971) antibodies were obtained from Cell Signaling Technology, Inc.

Techniques: Activation Assay, Protein-Protein interactions, Expressing, Western Blot, Control

Journal: International Journal of Molecular Medicine

Article Title: TGIF1 plays a carcinogenic role in esophageal squamous cell carcinoma through the Wnt/β-catenin and Akt/mTOR signaling pathways

doi: 10.3892/ijmm.2021.4910

Figure Lengend Snippet: TGIF1 promotes the activation of the Wnt/β-catenin and Akt/mTOR signaling pathways in vivo . Expression levels of Wnt/β-catenin-related and Akt/mTOR-related proteins were detected by immunohistochemistry in the KYSE-150 and Eca-109 tumors. TGIF1, TGFB induced factor homeobox 1; mTOR, mammalian target of rapamycin.

Article Snippet: Anti-Bcl-2 (cat. no. 12789-1-AP), anti-Bax (cat. no. 50599-2-Ig), anti-cyclin 1 (cat. no. 60186-1-Ig) and anti-GAPDH (cat. no. 60004-1-lg) antibodies were obtained from Proteintech Group, Inc. Anti-cleaved caspase-3 (cat. no. 9661), anti-Wnt3a (cat. no. 2391), anti-β-catenin (cat. no. 9562), anti-Akt (cat. no. 9272), anti-p-Akt (cat. no. 9271), anti-mTOR (cat. no. 2972) and anti-p-mTOR (cat. no. 2971) antibodies were obtained from Cell Signaling Technology, Inc.

Techniques: Activation Assay, Protein-Protein interactions, In Vivo, Expressing, Immunohistochemistry

Journal: Molecular Medicine Reports

Article Title: Sanggenon C protects against cardiomyocyte hypoxia injury by increasing autophagy

doi: 10.3892/mmr.2017.7646

Figure Lengend Snippet: Effect of Sanggenon C on AMPKα/mTOR/FOXO3a signaling. (A) Western blot analysis the effect of Sanggenon C (100 µM) on the activation of AMPKα pathways including phosphorylated (p-) and total (T-) AMPKα, mTOR and FOXO3a with (B) densitometry analysis (n=6). (C) TUNEL staining of cardiomyocytes pretreated with Sanggenon C (100 µM) or AMPKa inhibitor, CpC (20 µM) and exposed to hypoxia for 24 h (n=3) and (D) quantification of TUNEL-positive cells. (E) Western blot analysis the effect of Sanggenon C (100 µM) and CpC on the transformation of LC3 I to LC3 II (n=6). *P<0.05 vs. normoxia-PBS; # P<0.05 vs. hypoxia-PBS. HD, high dose 100 µM Sanggenon C; p-, phospho; T-, total; AMPKα, AMP-activated protein kinase α; mTOR, mechanistic target of rapamycin; FOXO3a, forkhead box O3a; TUNEL, terminal deoxynucleotidyl-transferase-mediated dUTP nick-end labeling; LC3, microtubule-associated proteins 1A/1B light chain 3; CpC, compound C.

Article Snippet: The primary antibodies included phosphorylated AMP-activated protein kinase α (p-AMPKα; cat. no. 2535), total (T)-AMPKα (cat. no. 2603P), p-mechanistic target of rapamycin (mTOR; cat. no. 2971), T-mTOR (cat. no. 2983), p-forkhead box O3a (FOXO3a; cat. no. 9465P), T-FOXO3a (cat. no. 2497P), Bcl-2 associated X apoptosis regulator (Bax; cat. no. 2722), Bcl-2 apoptosis regulator (Bcl-2; cat. no. 2870), and GAPDH (cat. no. 2118; all purchased from Cell Signaling Technology, Inc., Danvers, MA, USA).

Techniques: Western Blot, Activation Assay, TUNEL Assay, Staining, Transformation Assay, End Labeling