Journal: Molecules

Article Title: Oridonin Induces Apoptosis in Esophageal Squamous Cell Carcinoma by Inhibiting Cytoskeletal Protein LASP1 and PDLIM1

doi: 10.3390/molecules28020805

Figure Lengend Snippet: Top Candidate Proteins Involved in Oridonin-treated ESCC.

Article Snippet: Membranes were blocked with 5% Beef Serum Albumin (BSA) and then incubated with primary antibodies: mouse Hsp70 antibody (mAb; Abcam, Boston, MA, USA), rabbit ENO1, LASP1, GAPDH antibodies (#3810 (pAb), #8636 (pAb), #2218 (mAb); cell signaling, Danvers, MA, USA), rabbit PDLIM1 antibody (ABclonal, Cambridge, MA, USA), and mouse β -Actin antibody (Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA).

Techniques: Variant Assay

Journal: Molecules

Article Title: Oridonin Induces Apoptosis in Esophageal Squamous Cell Carcinoma by Inhibiting Cytoskeletal Protein LASP1 and PDLIM1

doi: 10.3390/molecules28020805

Figure Lengend Snippet: Gene Set Enrichment Analysis and western blotting assessed that oridonin inhibits LASP1 and PDLIM1 on ESCC. ( A ) Molecular function classification of the protein candidates involved in oridonin treatment. The top three are cadherin binding, cadherin binding involved in cell–cell adhesion, and heat shock protein binding. ( B ) The top three cellular component classifications are focal adhesion, cell-substrate junction, and cell cortex. ( C ) The top three biological processes are negative regulation of apoptotic signaling pathway, removal of superoxide radicals, and cellular response to oxygen radicals. ( D ) Protein expression of LASP1 and PDLIM1 under oridonin treatment decreased compared to untreated cell lysates. Hsp70 expression increased in oridonin-treated cell lysates compared to untreated while ENO1 decreased a little in treated TE-8.

Article Snippet: Membranes were blocked with 5% Beef Serum Albumin (BSA) and then incubated with primary antibodies: mouse Hsp70 antibody (mAb; Abcam, Boston, MA, USA), rabbit ENO1, LASP1, GAPDH antibodies (#3810 (pAb), #8636 (pAb), #2218 (mAb); cell signaling, Danvers, MA, USA), rabbit PDLIM1 antibody (ABclonal, Cambridge, MA, USA), and mouse β -Actin antibody (Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA).

Techniques: Western Blot, Binding Assay, Protein Binding, Expressing

Journal: Molecules

Article Title: Oridonin Induces Apoptosis in Esophageal Squamous Cell Carcinoma by Inhibiting Cytoskeletal Protein LASP1 and PDLIM1

doi: 10.3390/molecules28020805

Figure Lengend Snippet: Interested Genes for Protein Assessment Involved in Oridonin Treatment.

Article Snippet: Membranes were blocked with 5% Beef Serum Albumin (BSA) and then incubated with primary antibodies: mouse Hsp70 antibody (mAb; Abcam, Boston, MA, USA), rabbit ENO1, LASP1, GAPDH antibodies (#3810 (pAb), #8636 (pAb), #2218 (mAb); cell signaling, Danvers, MA, USA), rabbit PDLIM1 antibody (ABclonal, Cambridge, MA, USA), and mouse β -Actin antibody (Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA).

Techniques: Binding Assay, Protein Binding

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Mediation of PKM2-dependent glycolytic and non-glycolytic pathways by ENO2 in head and neck cancer development

doi: 10.1186/s13046-022-02574-0

Figure Lengend Snippet: AP-III-a4 inhibits cell proliferation and glucose metabolism in HNSCC cells through suppressing ENO2-mediated downstream signaling. a Effect of AP-III-a4 treatment on ENO1 and ENO2 protein levels determined by western blotting. UM-1 and Cal27 cells were treated with AP-III-a4 at a dose range (0, 1, 2.5, 5µM) for 48 h. b - e Effect of AP-III-a4 treatment on ENO2-mediated downstream targets ( b ), cell proliferation ( c ), and intracellular levels of ATP ( d ) and glucose uptake ( e ). UM-1 and Cal27 cells were treated with or without 5µM AP-III-a4 for 48 h. * p <0.05; ** p <0.01

Article Snippet: ERK1/2, p-ERK1/2, AKT, p-AKT, P70S6K, p-P70S6K, cyclin D1 (CCND1), cyclin B, cyclin E, CDK2, CDK4, CDK6 and ENO1 antibodies were purchased from Cell Signaling Technology.

Techniques: Western Blot

Journal: PLoS ONE

Article Title: Systematic Analysis Reveals Elongation Factor 2 and α-Enolase as Novel Interaction Partners of AKT2

doi: 10.1371/journal.pone.0066045

Figure Lengend Snippet: A: Activity of immuno-captured α-enolase (ENO1) from HEK293T cells after stimulation with insulin and AKT-inhibition with CCT128930. Data represent means ± SD of n = 7 experiments. B: Detection of immuno-captured α-enolase after α-enolase activity assay. After detection of α-enolase blots were incubated with anti-pan AKT antibody. Note that AKT was also detected after isolation of α-enolase by the α-enolase antibody.

Article Snippet: Primary Antibodies (Cell Signaling Technology, abcam): AKT2 Mouse mAB (5239), eEF2-Antibody (2332), GAPDH Rabbit mAB (2118), AKT1 Mouse mAB (2967), anti-ENO1-antibody (ab85086).

Techniques: Activity Assay, Inhibition, Incubation, Isolation