Journal: Cell Death and Differentiation

Article Title: Deubiquitinating enzyme USP33 restrains docetaxel-induced apoptosis via stabilising the phosphatase DUSP1 in prostate cancer

doi: 10.1038/s41418-019-0473-8

Figure Lengend Snippet: a C4-2B and PC3 cells were transfected with control (Ctrl) or USP33 siRNA (USP33 RNAi #1 and #2, 20 nM) for 48 h and then treated with DMSO or docetaxel (Doc) (10 nM) for 24 h as indicated. The indicated molecules were examined by western blot assay. b USP33+/+ and USP33−/− PC3 cells were transfected with Mock, WT-USP33 or Mut-USP33 vector as indicated for 48 h and then treated with DMSO or docetaxel (Doc) (10 nM) for 24 h as indicated. The indicated molecules were examined by western blot assay. c USP33+/+ and USP33−/− PC3 cells were treated with DMSO or docetaxel (Doc) (10 nM) for 24 h in the presence of SP600125 (20 μM), SB203580 (20 μM) or transfected with JNK1/2 siRNA (20 nM) as indicated, and the indicated molecules were examined by western blot assay. One representative experiment of three is shown in a to c. Similar results were obtained in three independent experiments. d USP33+/+ and USP33−/− PC3 cells treated as in c were incubated with DMSO or docetaxel (10 nM) for 48 h. Annexin V+ apoptotic cells were quantified by Annexin V/PI assay. Error bars represent the mean ± s.d. of triplicate samples derived from one representative experiment. Similar results were obtained in three independent experiments. ***P < 0.001; ns, not significant (one-way ANOVA followed by Bonferroni multiple comparison using PRISM software).

Article Snippet: Antibodies specific for IkBa (44D4, #4812), phospho-ERK1/2 (Thr202/Tyr204) (197G2, #4377), JNK (#9252), phospho-JNK1/2 (Thr183/Tyr185) (81E11, #4668), p38 MAPK (D13E1, #8690), phospho-p38 (Thr180/Tyr182) (12F8, #4631), phospho-AKT (Thr308, #9275), cleaved caspase 3 (Asp175) (5A1E, #9664), cleaved PARP (Asp214) (D64E10, #5625), ubiquitin (P4D1, #3936), K48-linkage specific polyubiquitin (#4289) and Myc-tag (9B11, Sepharose bead conjugate, #3400) were purchased from Cell Signaling Technology (Beverly, MA, USA).

Techniques: Transfection, Western Blot, Plasmid Preparation, Incubation, Derivative Assay, Software

Journal: International Journal of Molecular Sciences

Article Title: Somatostatin, Cortistatin and Their Receptors Exert Antitumor Actions in Androgen-Independent Prostate Cancer Cells: Critical Role of Endogenous Cortistatin

doi: 10.3390/ijms232113003

Figure Lengend Snippet: Molecular consequences of somatostatin (SST) or cortistatin (CORT) treatment in androgen-independent (AI) prostate cancer (PCa) cells. ( A , B ) Phosphorylation levels of protein belonging to different oncogenic signaling pathways (AKT, ERK, JNK, PTEN and AR) in response to SST and CORT treatment in AI-PCa cells. Phospho-protein levels were normalized by the total amount of each respective protein. Protein data were represented as percent of vehicle-treated cells (set at 100%). ( C ) Fold change in markers of proliferation, migration, and PCa-aggressiveness in response to SST and CORT treatment in AI-PCa cells. Gene expression was represented as the percentage of vehicle-treated cells (set at 100%). Asterisks (* p < 0.05; ** p < 0.01 and *** p < 0.001) indicate statistically significant differences between treatment and vehicle-treated cells. N.D: Non-detected. Ctrl: Control.

Article Snippet: Membranes were blocked with 5% non-fat dry milk in Tris-buffered saline/0.05% Tween-20 and incubated overnight with the specific primary antibodies at 1:1000 dilution [phospho-AKT (p-AKT; #4060S; Cell-Signaling, Barcelona, Spain), AKT (#9272S; Cell-Signaling, Barcelona, Spain), phospho-ERK (#4370S; Cell-Signaling), ERK (#9102S; Cell-Signaling), phospho-JNK (#AF1206; RD system), JNK (#AF1387; RD system), phospho-PTEN (#S380; Cell-Signaling), PTEN (9552S; Cell-Signaling), phospho-AR (#16969; Cell-Signaling), AR (ab133273; Abcam)].

Techniques: Migration, Expressing