Journal: Cancers

Article Title: Chondrosarcoma: Multi-Targeting Therapeutic Effects of Doxorubicin, BEZ235, and the Small Molecule Aspartyl-Asparaginyl-β-hydroxylase Inhibitor SMI1182

doi: 10.3390/cancers17101671

Figure Lengend Snippet: Effects of SMI1182 (SMI), DOX, and BEZ on the expression of mid-level components of the iInsulin/IGF1-Akt-mTOR pathway in CS1 cells. Magnetic bead-based protein and phosphoprotein 11-plex ELISAs measured frontal lobe levels of ( A ) Akt, ( B ) GSK-3α, ( C ) GSK-3β, ( D ) PTEN, ( E ) pS473-Akt, ( F ) pS21-GSK-3α, ( G ) pS9-GSK-3β, ( H ) pS380-PTEN, and the calculated levels of relative phosphorylation of ( I ) p/T-Akt, ( J ) p/T-GSK-3α, and ( K ) p/T-GSK-3β, and ( L ), and p/T-PTEN, (compared with total protein) in cells (n = 4/group) treated for 48 h with Veh, SMI, DOX, or BEZ. Graphed values correspond to arbitrary fluorescent light units (FLU). Inter-group comparisons were made using two-way ANOVA tests with post -hoc Tukey tests. Software-generated p -values corresponding to significant differences ( p ≤ 0.05) are shown in the panels.

Article Snippet: We used 11-Plex MILLIPLEX Akt/mTOR Total and Phosphoprotein Magnetic Bead Kits (MilliporeSigma, Burlington, MA, USA) to evaluate the effects of treatment on mTOR signaling mechanisms in CS cells ( ).

Techniques: Expressing, Phospho-proteomics, Software, Generated

Journal: Cancers

Article Title: Chondrosarcoma: Multi-Targeting Therapeutic Effects of Doxorubicin, BEZ235, and the Small Molecule Aspartyl-Asparaginyl-β-hydroxylase Inhibitor SMI1182

doi: 10.3390/cancers17101671

Figure Lengend Snippet: Effects of SMI1182 (SMI), DOX, and BEZ on the expression of mid-level components of the iInsulin/IGF1-Akt-mTOR pathway in CDS11 cells. Magnetic bead-based protein and phosphoprotein 11-plex ELISAs measured frontal lobe levels of ( A ) Akt, ( B ) GSK-3α, ( C ) GSK-3β, ( D ) PTEN, ( E ) pS473-Akt, ( F ) pS21-GSK-3α, ( G ) pS9-GSK-3β, ( H ) pS380-PTEN, and the calculated levels of relative phosphorylation of ( I ) p/T-Akt, ( J ) p/T-GSK-3α, and ( K ) p/T-GSK-3β, and ( L ), and p/T-PTEN, (compared with total protein) in cells (n = 4/group) treated for 48 h with Veh, SMI, DOX, or BEZ. Graphed values correspond to arbitrary fluorescent light units (FLU). Inter-group comparisons were made using two-way ANOVA tests with post -hoc Tukey tests. Software-generated p -values corresponding to significant differences ( p ≤ 0.05) are shown in the panels.

Article Snippet: We used 11-Plex MILLIPLEX Akt/mTOR Total and Phosphoprotein Magnetic Bead Kits (MilliporeSigma, Burlington, MA, USA) to evaluate the effects of treatment on mTOR signaling mechanisms in CS cells ( ).

Techniques: Expressing, Phospho-proteomics, Software, Generated

Journal: Cancers

Article Title: Chondrosarcoma: Multi-Targeting Therapeutic Effects of Doxorubicin, BEZ235, and the Small Molecule Aspartyl-Asparaginyl-β-hydroxylase Inhibitor SMI1182

doi: 10.3390/cancers17101671

Figure Lengend Snippet: Effects of SMI1182 (SMI), DOX, and BEZ on the expression of downstream signaling through mTOR in CS1 cells. Magnetic bead-based protein and phosphoprotein 11-plex ELISAs measured immunoreactivity to ( A ) TSC2, ( B ) mTOR, ( C ) RPS6, ( D ) P70S6K, ( E ) pS939-TSC2, ( F ) pS2448-mTOR, ( G ) pS235/236-RPS6, ( H ) pT412-P70S6K, and the calculated relative levels of signaling molecule phosphorylation of ( I ) TSC, ( J ) mTOR, ( K ) RPS6, and ( L ) P70S6K in cells (n = 4 cultures/group) treated for 48 h with Veh, SMI, DOX, or BEZ. Graphed values correspond to arbitrary fluorescent light units (FLU). Inter-group comparisons were made by a two-way ANOVA . Software-calculated p -values reflecting significant differences ( p ≤ 0.05) are shown in the panels.

Article Snippet: We used 11-Plex MILLIPLEX Akt/mTOR Total and Phosphoprotein Magnetic Bead Kits (MilliporeSigma, Burlington, MA, USA) to evaluate the effects of treatment on mTOR signaling mechanisms in CS cells ( ).

Techniques: Expressing, Phospho-proteomics, Software

Journal: Cancers

Article Title: Chondrosarcoma: Multi-Targeting Therapeutic Effects of Doxorubicin, BEZ235, and the Small Molecule Aspartyl-Asparaginyl-β-hydroxylase Inhibitor SMI1182

doi: 10.3390/cancers17101671

Figure Lengend Snippet: Effects of SMI1182 (SMI), DOX, and BEZ on the expression of downstream signaling through mTOR in CDS11 cells. Magnetic bead-based protein and phosphoprotein 11-plex ELISAs measured immunoreactivity to ( A ) TSC2, ( B ) mTOR, ( C ) RPS6, ( D ) P70S6K, ( E ) pS939-TSC2, ( F ) pS2448-mTOR, ( G ) pS235/236-RPS6, ( H ) pT412-P70S6K, and the calculated relative levels of signaling molecule phosphorylation of ( I ) TSC, ( J ) mTOR, ( K ) RPS6, and ( L ) P70S6K in cells (n = 4 cultures/group) treated for 48 h with Veh, SMI, DOX, or BEZ. Graphed values correspond to arbitrary fluorescent light units (FLU). Inter-group comparisons were made by a two-way ANOVA . Software-calculated p -values reflecting significant differences ( p ≤ 0.05) are shown in the panels.

Article Snippet: We used 11-Plex MILLIPLEX Akt/mTOR Total and Phosphoprotein Magnetic Bead Kits (MilliporeSigma, Burlington, MA, USA) to evaluate the effects of treatment on mTOR signaling mechanisms in CS cells ( ).

Techniques: Expressing, Phospho-proteomics, Software

Journal: Scientific reports

Article Title: Neuroglobin regulates autophagy through mTORC1/RAPTOR/ULK-1 pathway in human neuroblastoma cells.

doi: 10.1038/s41598-025-91701-w

Figure Lengend Snippet: Fig. 5. NGB overexpression regulates autophagy targeting mTOR signaling pathway. CTRL and stably transfected SH-SY5Y-NGB-FLAG cells were lysed in lysis buffer. The lysates were analyzed by western blot to detect P-Ser2448-mTOR or total mTOR protein levels, using anti-P-Ser2448-mTOR pAb or anti-total mTOR pAb or P-Ser757-ULK1 or total ULK1 using anti-P- Ser757-ULK1 pAb. Loading control was evaluated using mouse anti-Beta-Actin mAb. Results represent the mean ± SD from three independent experiments (n = 3). *** p < 0.001. Uncropped blot images are shown in Supplementary Information file.

Article Snippet: Membranes were blocked with 5% w/v non-fat dried milk (sc-2325, Santa Cruz Biotechnology, Dallas, Texas, USA) in Tris buffer saline (1706435, Bio-Rad Laboratories) (TBS), containing 0.05% v/v Tween 20 (1706531, Bio-Rad Laboratories) (TBST) and probed with rabbit anti-LC3 pAb (NB100-2331, Novus Biologicals, Centennial, Colorado, USA), rabbit anti-SQSTM1 mAb (Cell Signaling Technology, Danvers, Massachusetts, USA), rabbit anti-P-Ser757-ULK1 pAb (6888, Cell Signaling Technology), rabbit anti-P-Ser2448-mTOR pAb (2971, Cell Signaling Technology), rabbit total ULK1 mAb (8054, Cell Signaling Technology) or rabbit anti-total mTOR mAb (2983, Cell Signaling Technology).

Techniques: Over Expression, Stable Transfection, Transfection, Lysis, Western Blot, Control