Project description:SK-MEL-2 cells treated with Ad-E2F-1 (MOI 2), Ad-E2F-1 (MOI 2)plus doxorubicin 0.1uM, or Ad-LacZ (MOI 2) plus doxorubicin 0.1uM. The combination of E2F-1 gene therapy and chemotherapy produces a synergistic effect on melanoma cell apoptosis. However, the molecular mechanisms have not been fully elucidated. The purpose of this study was to identify novel genes or pathways that may play key roles in apoptosis when E2F-1 gene therapy is combined with doxorubicin chemotherapy. MATERIALS AND METHODS: SK-MEL-2 melanoma cells were infected with Ad-E2F-1 alone, Ad-E2F-1 plus doxorubicin, or Ad-LacZ plus doxorubicin. After 16 hours of treatment, the total RNA was extracted from these cells and subjected to microarray analysis. Quantitative real-time PCR was performed to confirm the microarray data. RESULTS: Our results showed that the combination treatment of Ad-E2F-1 and doxorubicin affected the expression of cytokines, transcription factors, as well as genes involved in signal transduction, cell cycle regulation and apoptosis. CONCLUSION: Our findings have identified, for the first time, novel molecular targets and pathways that led to apoptosis in melanoma cells when Ad-E2F-1 was combined with doxorubicin. The molecular information provided here will enhance further mechanistic studies.
Project description:Human serum derived macrophages were infected with Mtb expressing a transcriptional reporter of viability and on day 5, the cells were sorted for RFP+GFP+ macrophages or RFP+GFP- macrophages.
Project description:ES derived Flk-1+ cells were separated by sorting into Hoxb6 positive and negative populations by RFP Hoxb6 reporter. Gene expression was compared between these two groups. Duplicate analysis of Hoxb6-RFP positive and negative Flk-1+ cells.
Project description:ES derived Flk-1+ cells were separated by sorting into Hoxb6 positive and negative populations by RFP Hoxb6 reporter. Gene expression was compared between these two groups.
Project description:C57B6J mice carrying a LacZ allele at the Math1 locus (Math1-LacZ/+) were backcrossed at least 10 generations. The LacZ allele phenocopies a null allele. A total of 5 heterozygous and 5 homozygous null spinal cords were collected (staged at embryonic day 12.5) and individually processed (i.e. not pooled). RNA was extracted and prepared for hybridization to Affymetrix MOE430v2.0 chips under standard conditions (www.affymetrix.com)
Project description:We examined the impact of IL-34 on repopulation of Langerhans cells after inflammation Langerhans cells were FACS-purified from Il34+/LacZ and Il34LacZ/LacZ mice after UV treatment
