Journal: Gynecologic oncology

Article Title: MEK inhibition overcomes resistance to EphA2-targeted therapy in uterine cancer.

doi: 10.1016/j.ygyno.2021.08.003

Figure Lengend Snippet: Fig. 4. Effects of dasatinib, trametinib, and dasatinib plus trametinib on MAPK pathway and downstream targets. (A) Heatmap of reverse phase protein array (RPPA) analysis of dasatinib- resistant HEC-1A, Ishikawa, and RL-95 cells before and after treatment with dasatinib alone, trametinib alone, or dasatinib plus trametinib for 16 h. Data represent the means of triplicate measurements. (B) RPPA analysis of the normalized expression levels of pMAPK, BIM, JAG1, and c-MYC in dasatinib-resistant HEC-1A, Ishikawa, and RL-95 cells before and after treatment. Data represent the means of triplicate measurements. (C) Pathway analysis of target modulation in dasatinib-resistant RL-95 cells before and after treatment with dasatinib alone or with dasatinib plus trametinib, as detected by NetWalker pathway analysis. (D) Western blot analysis of BIM, c-MYC, cleaved PARP, HES1 and HEYL in RL-95 cells treated with 100 nM dasatinib alone, 1 μM ALWII alone, 100 nM trametinib alone, dasatinib plus trametinib, or ALWII plus trametinib for 48 h. β-actin was used as a loading control.

Article Snippet: Antibodies against p-MEK (Ser217/Ser221), #3958; p-p44/42 MAPK (Thr202/Tyr204), #9101; c-MYC (#5605); and cleaved PARP (#5625) were obtained from Cell Signaling Technology (Danvers, MA).

Techniques: Protein Array, Expressing, Western Blot, Control

Journal: Gynecologic oncology

Article Title: Entinostat, a selective HDAC1/2 inhibitor, potentiates the effects of olaparib in homologous recombination proficient ovarian cancer.

doi: 10.1016/j.ygyno.2021.04.015

Figure Lengend Snippet: Fig. 4. Olaparib combined with entinostat decreases proliferation, alters HR machinery and increases apoptosis in HR-proficient SKOV-3 cells. (A). Immunofluorescence showing that Ola + Ent significantly reduces BRCA1 and Ki-67 expression levels in SKOV-3 cells. DAPI was used as nuclear stain (40 × 100 μm scale bar). Quantification of BRCA1 and Ki-67 indicated significant downregulation in cells treated with Ola + Ent (*p < 0.05, **p < 0.005 Students t-test). (B) BRCA1 transcript levels showed significant downregulation in all treatment groups, with enhanced downregulation in Ola + Ent (*p < 0.012; **p < 0.0036; ***p < 0.0005, one-way ANOVA; Sidak's multiple comparison test) indicating that BRCA1 expression is affected markedly at the mRNA levels compared to protein levels. (C) Western blot analysis of SKOV-3 cells treated with Ola + Ent as described for SRB assay showing increased cleaved PARP expression in olaparib and Ola + Ent treated cells. Quantification showed significant upregulation of cleaved PARP, a marker of cell death (*p < 0.05; **p < 0.005, Students t-test). The antibody used was specific for fragments of PARP1. [Con – Control; Ola – Olaparib; Ent – Entinostat].

Article Snippet: The primary antibody was a rabbit polyclonal against cleaved PARP that detects PARP1 fragments, but not intact PARP1 (Cell Signaling Technology; 5625; 1:100) for overnight incubation at 4 °C and amousemonoclonal against β-actin (Life Technologies; MA1–140) was used as loading control.

Techniques: Expressing, Staining, Comparison, Western Blot, Sulforhodamine B Assay, Marker, Control

Journal: Gynecologic oncology

Article Title: Entinostat, a selective HDAC1/2 inhibitor, potentiates the effects of olaparib in homologous recombination proficient ovarian cancer.

doi: 10.1016/j.ygyno.2021.04.015

Figure Lengend Snippet: Fig. 6. Graphical summary indicating the mechanism for entinostat-induced olaparib sensitivity in HR-proficient cells. As described, entinostat causes breakdown of HR-DNA repair pathway, whereas olaparib inactivates PARP, crippling the cell's potential to repair SSBs. This in turn leads to accumulation of DSBs, which the cell is unable to repair due to defective HR repair pathway, accumulating unrepaired DNA ultimately resulting in cell death.

Article Snippet: The primary antibody was a rabbit polyclonal against cleaved PARP that detects PARP1 fragments, but not intact PARP1 (Cell Signaling Technology; 5625; 1:100) for overnight incubation at 4 °C and amousemonoclonal against β-actin (Life Technologies; MA1–140) was used as loading control.

Techniques:

Journal: International journal of molecular sciences

Article Title: Fibrinogen-Like Protein 1 Modulates Sorafenib Resistance in Human Hepatocellular Carcinoma Cells.

doi: 10.3390/ijms22105330

Figure Lengend Snippet: Figure 3. Effect of sorafenib on cell death and cell proliferation factors in high FGL1- and low FGL1-expressing hepatocellular carcinoma (HCC) cell lines. (A): Protein expression was analyzed by western blotting 48 h after treatment with 10 µM sorafenib. Expression levels of PCNA, p-ERK/ERK, LC3-II, and cleaved PARP1 quantified in Huh7 (B), Hep3B (C), SNU387 (D), and SNU475 (E) cell lines. Data are presented as the mean ± standard deviation (SD) (n = 3). * p < 0.05, ** p < 0.01, and *** p < 0.001, significantly different from 0 µM sorafenib-treated cells.

Article Snippet: 2021, 22, 5330 10 of 12 dilution; Cell Signaling Technologies), anti-LC3-II (1:3000 dilution; Novus Biologicals, Littleton, CO, USA), anti-cleaved PARP1 (1:1000 dilution; Cell Signaling Technologies), anti-Beclin-1 (1:1000 dilution; Cell Signaling Technologies), and anti-β-actin (1:2000 dilution; Sigma-Aldrich, St. Louis, MO, USA).

Techniques: Expressing, Western Blot, Standard Deviation