Journal: Human reproduction (Oxford, England)

Article Title: Dormancy and activation of human oocytes from primordial and primary follicles: molecular clues to oocyte regulation.

doi: 10.1093/humrep/dex238

Figure Lengend Snippet: Figure 6 Intraovarian distribution of VASA, FOXO1 and EIF4E in human oocytes from primordial and primary follicles. (A) VASA localized to oocytes in both primordial and primary follicles and presented here is an example of VASA in primordial oocytes. DAPI staining identifies the nucleus of cells in this slide and the surrounding flat granulosa cells are characteristic for oocytes from primordial follicles. (B) FOXO1 localizes intensively in the nucleus of the oocyte from primordial follicles, as expected. Hoechst staining identifies the nucleus of cells in this slide and the surrounding flat gran- ulosa cells are characteristic for oocytes from primordial follicles. (C) FOXO1 is less intense and less nucleus-associated in oocytes from primary folli- cles. Hoechst staining identifies the nucleus of cells in this slide and the surrounding cuboidal granulosa cells are characteristic for oocytes from primary oocytes. (D) EIF4E is less intense in the primordial oocytes as expected. Hoechst staining identifies the nucleus of cells in this slide and the surrounding flat granulosa cells are characteristic for oocytes from primordial follicles. (E) EIF4E is more intense in the oocyte from the primary follicle. Hoechst staining identifies the nucleus of cells in this slide and the surrounding cuboidal granulosa cells are characteristic for oocytes from primary follicles. Scale bars: 30 μm.

Article Snippet: Rehydration and antigen retrieval was performed as described elsewhere (Stubbs et al., 2005) followed by serum block (30 min), then primary antibody: (1/200) anti-VASA Rabbit mAb (Abcam, Cambridge, U.K.), (1/100) anti-FOXO1 Rabbit mAb (Cell Signalling Technology, Danvers, MA, USA), or (1/1600) anti-EIF4E Rabbit mAb (Cell Signalling Technology, Danvers, MA, USA), overnight at 4°C.

Techniques: Staining

Journal: Endocrine-Related Cancer

Article Title: Arsenic trioxide synergizes with everolimus (Rad001) to induce cytotoxicity of ovarian cancer cells through increased autophagy and apoptosis

doi: 10.1530/erc-12-0150

Figure Lengend Snippet: Figure 4 Combination of ATO and Rad001 induces apoptosis and autophagy in SKOV3 and OV2008 ovarian cancer cells. Combination treatment increased the levels of Atg5–Atg12 (A) and decreased the levels of p-AKT, p-4E-BP1 compared with Rad001 or ATO alone (B). Rad001 treatment decreased expression levels of p-p70S6K and p-4E-BP1. In SKOV3 cell line, ATO could induce the express of p-p70S6K protein level. p-eIF4E protein level did not change significantly in response to the combination treatment in both cells. (C) Beclin1 (Atg6) level did not change in response to the combination treatment. (D) p-ERK1 protein level decreased more pronouncedly in response to the combination treatment than single-drug treatment. The p-JNK protein level was similar in different treatment groups.

Article Snippet: Fetal bovine serum (FBS), RPMI 1640 medium, DMEM, sodium pyruvate, L-glutamine, penicillin, and streptomycin were purchased from Hyclone (Logan, UT, USA); ATO and Rad001 were from Sigma; CellTiter Glo assay was from Promega; Caspase-3/7 Assay Kit was from Anaspec 712 (Fremont, CA, USA); Annexin V apoptosis detection Kit was from eBioscience (San Diego, CA, USA); Lipofectamine 2000 transfection reagent was from Invitrogen (New York, NY, USA); rabbit anti-LC-3 polyclonal antibody was from GenScript (Piscataway, NJ, USA); rabbit anti-Beclin1, rabbit anti-cleaved caspase-3, and rabbit anti-PARP1 polyclonal antibodies, antibodies recognizing p70S6K, phosphop70S6K (Thr389), AKT, phospho-AKT (Ser473), 4E-BP1, p-4E-BP1 (Thr37/46), eIF-4E, p-eIF-4E (Ser209), c-Jun-NH2-terminal kinase (JNK), and p-JNK were from Cell Signaling (Danvers, MA, USA); rabbit anti-Atg5 monoclonal antibody was from Epitomics (Burlingame, CA, USA); rabbit polyclonal antibody to SQSTM1/p62 (sequestosome 1) was from GeneTeX (Irvine, CA, USA); and mouse anti-b-actin antibody was from Sigma.

Techniques: Expressing