Journal: International Journal of Molecular Sciences

Article Title: Alpha Ketoglutarate Exerts In Vitro Anti-Osteosarcoma Effects through Inhibition of Cell Proliferation, Induction of Apoptosis via the JNK and Caspase 9-Dependent Mechanism, and Suppression of TGF-β and VEGF Production and Metastatic Potential of Cells

doi: 10.3390/ijms21249406

Figure Lengend Snippet: Effect of AKG on phosphorylation of MAP kinases and the influence of the selective JNK inhibitor (SP600125) on AKG-induced apoptosis in Saos-2 cells. The cells were incubated without or with AKG for 6 h and 24 h, and phosphorylated and total ERK1/2, JNK and p38 levels were determined with the ELISA assay. Quantification of the amounts of phosphorylated to total MAP kinases ( A – C ). The cells were treated with 50 mM AKG without or with SP600125 (5 μM) and harvested after 72 h of treatment for apoptosis analysis. The representative dot plots indicate the percentage of An − /PI + necrotic cells (Q1), An + /PI + late apoptotic cells (Q2), An − /PI − viable cells (Q3), and An + /PI − early apoptotic cells (Q4) in the AKG or/and SP600125-treated Saos-2 cell cultures ( D ). Quantification of the amounts of phosphorylated to total JKN kinase ( E ) and histogram representation of the quantitative percentage of apoptotic (early + late apoptosis) cells ( F ) in the control, SP600125, AKG, and SP600125 + AKG-treated Saos-2 cell cultures. Data are expressed as means ± SD for three independent experiments. ** p < 0.01, *** p < 0.001 in comparison to the control; one-way ANOVA test.

Article Snippet: The quantification of the intracellular levels of total and phosphorylated JNK, ERK1/2, p38, and AKT kinases and the contents of cyclin D1 and p21 proteins in the treated cells was carried out using the PathScan ® ELISA kits: Total SAPK/JNK Sandwich ELISA Kit, Phospho-SAPK/JNK (Thr183/Tyr185) Sandwich ELISA Kit, Total p44/42 MAPK (Erk1/2) Sandwich ELISA Kit, Phospho-p44/42 MAPK (Thr202/Tyr204) Sandwich ELISA Kit, Phospho-p38 MAPK (Thr180/Tyr182) Sandwich ELISA Kit, Total Cyclin D1 Sandwich ELISA Kit, Total p21 Waf1/Cip1 Sandwich ELISA Kit (Cell Signaling Technology Danvers, MA, USA), and p38 MAPK alpha ELISA Kit (Abcam, Cambridge, UK) according to the manufacturer’s instructions as described previously [ ].

Techniques: Incubation, Enzyme-linked Immunosorbent Assay

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Identification of a novel HLA-A2-restricted mutated Survivin epitope and induction of specific anti-HCC CTLs that could effectively cross-recognize wild-type Survivin antigen

doi: 10.1007/s00262-012-1323-4

Figure Lengend Snippet: Survivin candidate epitope predicted by computational methods

Article Snippet: The ascites supernatant were used to detect the Survivin levels with the PathScan Survivin Sandwich ELISA Kit(Cell Signaling Technology, Danvers, MA)according to the manufacturer’s directions.

Techniques: Sequencing

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Identification of a novel HLA-A2-restricted mutated Survivin epitope and induction of specific anti-HCC CTLs that could effectively cross-recognize wild-type Survivin antigen

doi: 10.1007/s00262-012-1323-4

Figure Lengend Snippet: Specific lysis of CTLs generated from different Survivin-derived peptides against peptide-pulsed T2 cells. Immune effector cells were incubated with target cells at the effector/target ratios shown. CTL assays were performed following a 4-h incubation period. E represents peptide-induced CTLs effectors and T represents peptide-pulsed T2 target cells. Cytotoxic T lymphocytes generated from HIVpol476, served as a NP. Data points represent the mean ± SD as measured by quantitation of LDH release. **P < 0.01 compared to the NP group

Article Snippet: The ascites supernatant were used to detect the Survivin levels with the PathScan Survivin Sandwich ELISA Kit(Cell Signaling Technology, Danvers, MA)according to the manufacturer’s directions.

Techniques: Lysis, Generated, Derivative Assay, Incubation, Quantitation Assay