Journal: Nature cell biology

Article Title: A complex secretory program orchestrated by the inflammasome controls paracrine senescence

doi: 10.1038/ncb2784

Figure Lengend Snippet: (a) Co-culture with cells undergoing OIS induces the arrest of normal IMR90 cells. Normal IMR90 human fibroblasts expressing the fluorescent marker mCherry (IMR90 Cherry), were mixed with IMR90 ER:RAS cells. When indicated 200 nM 4OHT was added to activate ER:RAS. Growth curves represent the number of IMR90 ER:RAS (left) or IMR90 Cherry cells (right) present in the co-cultures. Data is a representative experiment of n=2 independent experiments for days 0-7 and mean +/− s.d. of n= 3 independent experiments for day 8. The source data for 2 independent experiments and statistics for day 8 (Student’s t-test) are provided in . (b) Co-culture with IMR90 MEK:ER cells induces the arrest of normal IMR90 cells. The percentage of positive BrdU cells for each population in the cocultures 4 days after 4OHT induction is shown. Data is a representative experiment. The source data for 2 independent experiments are provided in . (c) IMR90 ER:RAS or IMR90 ER cells were co-cultured with normal IMR90 Cherry fibroblasts. BrdU incorporation at day 7 shows that co-culture with IMR90 ER:RAS but not with IMR90 ER induces the arrest of normal IMR90 Cherry cells (centre). Data is a representative experiment. The source data for 2 independent experiments are provided in . Representative images are shown (right). Scale bar, 30 μm. (d) Normal HMEC or IMR90 cells suffer growth arrest when co-cultured with HMEC cells undergoing OIS. HMEC-hTERT (centre) or IMR90 (right) cells expressing Cherry as a fluorescent marker were co-cultured with HMEC-TERT ER:RAS or HMEC-TERT vector cells in the presence of 100 nM 4OHT. Growth curves showing growth arrest of HMEC-TERT Cherry (centre) or IMR90-Cherry cells (right) are shown. Data is a representative experiment.

Article Snippet: HMEC-hTert cells were cultured in mammary epithelial cell growth media (PromoCell).

Techniques: Co-Culture Assay, Expressing, Marker, Cell Culture, BrdU Incorporation Assay, Plasmid Preparation

Journal: Oncotarget

Article Title: Inhibiting NFAT1 for breast cancer therapy: New insights into the mechanism of action of MDM2 inhibitor JapA

doi:

Figure Lengend Snippet: A. The chemical structure of JapA. B. Human breast epithelial cells and breast cancer cells were exposed to 2 μM JapA for 24 h. The protein level of NFAT1 was detected by western blotting. C. MCF-7 and MDA-MB-231 cells were exposed to various concentrations of JapA for 24 h for the protein expression of NFAT1, c-Myc and COX-2. The experiments were repeated three times. D, E. JapA was administered by i.p. injection (15 or 30 mg/kg/d, 5 d/wk) to nude mice bearing MCF-7 or MDA-MB-231 xenograft tumors. After 5 weeks (MCF-7) or 3 weeks (MDA-MB-231) JapA treatment, the tumor tissues were analyzed for (D) the protein expression of NFAT1 by immunohistochemistry (scale bar, 20 μm) and (E) the protein expression of NFAT1, c-Myc and COX-2 by Western blotting (each lane represents a different tumor sample). The intensity ratio under each band was obtained by IMAGEJ software analysis normalized on untreated control.

Article Snippet: HMLE cells were grown in mammary luminal epithelial cell growth medium (Zen-Bio, Inc.).

Techniques: Western Blot, Expressing, Injection, Immunohistochemistry, Software, Control