Comparative analysis of gene expression changes upon Cabut or dE2F1/dDP overexpression in the Drosophila wing
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ABSTRACT: Investigation of whole genome gene expression level changes in dissected Drosophila wings at the wandering L3 larval stage, 24h after pupa formation and 36h after pupa formation, expressing either UAS-Cabut or UAS-dE2F1 + UAS-dDP under control of apterous-gal4 with a tubulin driven temperature-sensitive gal80 transgene, compared to the parental strain lacking UAS transgenes.
Project description:Investigation of whole genome gene expression level changes in dissected Drosophila wings at the wandering L3 larval stage, 24h after pupa formation and 36h after pupa formation, expressing either UAS-Cabut or UAS-dE2F1 + UAS-dDP under control of apterous-gal4 with a tubulin driven temperature-sensitive gal80 transgene, compared to the parental strain lacking UAS transgenes. A 36-chip study using total RNA recovered from four independently isolated samples of 10 dissected Drosophila wings at the indicated developmental stages expressing the indicated UAS-transgenes or the non-UAS-containing matched control strain. One sample (2D) did not pass our internal quality control and, therefore, was not included in the analysis. The dE2F1+dDP data included here is further discussed in L. Buttitta, A.J. Katzaroff, B.A. Edgar (2010). A robust cell cycle control mechanism limits E2F-induced proliferation of terminally differentiated cells in vivo. Journal of Cell Biology vol. 189, 981-96 (PMID 20548101). The Cabut data included here is further discussed in A.J. Katzaroff et al. (2011). The Krüppel-like-factor cabut has cell cycle regulatory properties similar to E2F.
Project description:Investigation of whole genome gene expression changes in dissected Drosophila wings at the wandering L3 larval stage and 24h after pupa formation, expressing UAS-activated Thickveins Q-D under control of apterous-gal4 with a tubulin driven temperature sensitive gal80 transgene, compared to the parental strain lacking the UAS-Thickveins transgene
Project description:Investigation of whole genome gene expression changes in dissected Drosophila wings at the wandering L3 larval stage and 24h after pupa formation, expressing UAS-activated Thickveins Q-D under control of apterous-gal4 with a tubulin driven temperature sensitive gal80 transgene, compared to the parental strain lacking the UAS-Thickveins transgene A 20 chip study using total RNA recovered from five independently isolated samples of 10 dissected Drosophila wings at the indicated developmental stages expressing UAS-activated thickveins or the non-UAS containing matched control strain.
Project description:Investigation of whole genome gene expression level changes in dissected Drosophila wings of the genotype w1118, at the wandering L3 larval stage, 2h, 6h, 18h, 24h and 36h After pupa formation. Stages after pupa formation were compared to the larval L3 stage to identify changes in gene expression. The data included here is further discussed in O’Keefe, Thomas, Edgar and Buttitta (2012). Combinatorial control of temporal gene expression in the Drosophila wing by enhancers and core promoters Submitted to BMC Genomics
Project description:Investigation of whole genome gene expression level changes in dissected Drosophila wings of the genotype w1118, at the wandering L3 larval stage, 2h, 6h, 18h, 24h and 36h After pupa formation. Stages after pupa formation were compared to the larval L3 stage to identify changes in gene expression. The data included here is further discussed in O’Keefe, Thomas, Edgar and Buttitta (2012). Combinatorial control of temporal gene expression in the Drosophila wing by enhancers and core promoters Submitted to BMC Genomics A 24 chip study using total RNA recovered from three to four independently isolated samples of 10 dissected Drosophila wings at the indicated developmental stages from flies with the genotype w1118;+;+ (Bloomington Stock #3605). For sample 36BD, total RNA from sample 36B and 36D were pooled, labeled and hybridized independently.
Project description:To investigate the difference of miRNA expression between lung cancer cell A549 and its DDP-resistant cell strain A549/DDP, we have employed miRNA microarray expression to discover the difference expression of miRNAs of A549 cells and A549/DDP. We conducted RT-qPCR to examine the expression levels of top differential expressed miRNAs, namely, miR-197-5p, miR-4443, miR-642a-3p, miR-27b-3p and miR-100-5p, confirming low variability between two methods. The A549/DDP was established from A549 in our laboratory, by exposing A549 to gradually increasing DDP concentrations, until the final concentration at 1μg/ml. To avoid the influence of drug to the A549/DDP cells, they were cultured in a drug-free medium for at least two weeks before gene expression analysis. miRNA expression of A549 and A549/DDP was then analzyed.
Project description:To investigate the difference of miRNA expression between lung cancer cell A549 and its DDP-resistant cell strain A549/DDP, we have employed miRNA microarray expression to discover the difference expression of miRNAs of A549 cells and A549/DDP. We conducted RT-qPCR to examine the expression levels of top differential expressed miRNAs, namely, miR-197-5p, miR-4443, miR-642a-3p, miR-27b-3p and miR-100-5p, confirming low variability between two methods.
Project description:To investigate the difference of miRNA expression in exosomes derived from A549 cells and its DDP-resistant cell strain A549/DDP, we have employed miRNA microarray expression to discover the difference expression of miRNAs in exosomes derived from A549 and A549/DDP. We conducted RT-qPCR to examine the expression levels of top differential expressed miRNAs, namely, miR-197-5p, miR-4443, miR-642a-3p, miR-27b-3p and miR-100-5p, confirming low variability between two methods. The A549/DDP was established from A549 in our laboratory, by exposing A549 to gradually increasing DDP concentrations, until the final concentration at 1μg/ml. To avoid the influence of drug to the A549/DDP cells, they were cultured in a drug-free medium for at least two weeks before gene expression analysis. After being incubated for 48-72h, the culture medium of cells was harvested. Exosomes were isolated by ultracentrifugation. And miRNA expression of exosomes derived from A549 and A549/DDP was then analzyed.
Project description:To investigate the difference of miRNA expression in exosomes derived from A549 cells and its DDP-resistant cell strain A549/DDP, we have employed miRNA microarray expression to discover the difference expression of miRNAs in exosomes derived from A549 and A549/DDP. We conducted RT-qPCR to examine the expression levels of top differential expressed miRNAs, namely, miR-197-5p, miR-4443, miR-642a-3p, miR-27b-3p and miR-100-5p, confirming low variability between two methods.
Project description:A 2-hour heat-shock at 37C was used to activate hs-FLP and an actin5C-FRT-stop-FRT-GAL4 transgene in larvae carrying any possible combination of the genetic elements UAS-Myc, UAS-Atu-IR, Max-/-. 48 hours later 16-27 wing imaginal discs were isolated from wandering L3 larvae and polyA-RNA was processed for sequencing.