Journal: Reproductive Sciences

Article Title: The Essential Role of GATA6 in the Activation of Estrogen Synthesis in Endometriosis

doi: 10.1177/1933719118756751

Figure Lengend Snippet: A, Differences in estradiol (E2) production in OSIS experimental groups. N = 4. Asterisk (*) indicates significantly lower concentration compared to other groups tested (P < .05). B, Differences in estradiol (E2) production in normal endometrial stromal cells (NoEMs) experimental groups. N = 4. Asterisk (*) indicates a significantly higher concentration compared to other groups tested (P < .05). C, Differences in estradiol (E2) production in skin fibroblast (BJ) experimental groups. N = 4.

Article Snippet: Human foreskin fibroblast cells (BJ cells), a nonimmortalized stromal cell line with a normal diploid karyotype, were obtained from ATCC and used as controls (cat. CRL-2522; Manassas, Virginia; n = 7).

Techniques: Concentration Assay

Journal: The EMBO Journal

Article Title: Asf1b, the necessary Asf1 isoform for proliferation, is predictive of outcome in breast cancer

doi: 10.1038/emboj.2010.335

Figure Lengend Snippet: Expression of Asf1b depends on the cycling status of cells. (A) (Left upper panel) Western blot analysis of total cell extracts from non-treated asynchronous (AS) and quiescent (G0) MCF7 breast cancer cells. Increasing amounts (X) of total cell extracts are loaded. We revealed Asf1a and Asf1b with a mix of the specific Asf1 antibodies (Supplementary Figure S1; Supplementary Table SI). We use CAF-1 p60 and cyclin A as markers for cell proliferation. α-Tubulin is a loading control. M: molecular weight marker. (Left lower panel) Asf1a and Asf1b mRNA levels in proliferating (AS) and quiescent (G0) MCF7 cells as determined by quantitative RT–PCR. We normalized levels to the reference gene RPLPO (de Cremoux et al, 2004) and set levels in proliferating cells to 100%. The error bars represent s.d. from two independent experiments. (Right panel) Specific expression of Asf1a, Asf1b and the largest subunit of CAF-1 (p150) revealed by immunofluorescence in MCF7 cells AS or G0. DAPI stains nuclei. Scale bar is 20 μm. (B) (Upper panel) Western blot analysis of total cell extracts from non-treated AS and G0 BJ primary foreskin fibroblasts and from primary IMR90 human diploid fibroblasts young (PD27), old (PD72) and senescent (PD80) performed as in A. (Lower panel) Asf1a and Asf1b mRNA levels in proliferating AS and G0 BJ primary fibroblasts or in young, old and senescent IMR90 human diploid primary fibroblasts as determined by quantitative RT–PCR performed as in A. The error bars represent s.d. from three and two independent experiments, respectively. (C) Flow cytometry analysis of the cell cycle distribution of MCF7 and BJ cells AS or G0.

Article Snippet: We used DMEM medium (GIBCO) for U-2-OS osteosarcoma (gift from J Bartek, Copenhagen), HeLa cervical carcinoma (gift from M Bornens, Paris), MCF7 and MDA-MB-231 breast adenocarcinoma cancer cell lines, MEMα medium (GIBCO) for BJ primary foreskin fibroblasts (CRL-2522, ATCC), RPMI medium (GIBCO) supplemented with 10 mg/ml insulin (Sigma) for Hs478T breast cancer cells (gift from O Delattre, Paris) ( Hackett et al, 1977 ), and DMEM medium (GIBCO) containing 30 ng/ml epidermal growth factor (TEBU) for Hs478Bst healthy mammary cells (ATCC) ( Hackett et al, 1977 ).

Techniques: Expressing, Western Blot, Control, Molecular Weight, Marker, Quantitative RT-PCR, Immunofluorescence, Flow Cytometry