Project description:The analysis of differentially regulated mRNAs candidates in THP-1 cells infected with H37Rv was conducted, comparing them to both uninfected THP-1 cells and transfected reference samples. THP-1 cells are monocytes that differentiate into macrophages after being treated with PMA for 48 hours. Total RNA was isolated from the infected and transfected THP-1 cells at 24 hours post-infection.
Project description:Differentially regulated miRNA candidates in H37Rv infected THP-1 cells were analysed with respect to uninfected THP-1 reference samples. THP-1 cells are monocytes differentiated to macrophages after treatment with PMA for 48 hrs. Total RNA was isolated from infected THP-1 cells after 24 hrs of infection, cDNA was synthesized for TLDA real time PCR reaction using TaqMan MicroRNA Reverse Transcription kit and Megaplex Human Pool A and Pool B stem loop RT primers (version 3.0) as per manufacturer’s protocol. Further real time reaction was performed on QuantStudio 12K Flex Real-Time PCR System (Applied Biosystems) by using cDNA (without pre-amplification) on TLDA card A and card B (version 3.0).
Project description:To compare gene expression changes induced by infection with Mycobacterium tuberculosis (Mtb) with changes induced by purified Mtb products, we infected THP-1 cells with Mtb strain H37Rv or treated with purified Mtb products, then performed RNAseq.
Project description:Perturbation of host physiology by intracellular Mycobacterium tuberculosis (H37Rv) can be reflected by changes in gene expression pattern of host genes. Total RNA was isolated from PMA differentiated uninfected cells or cells infected with H37Rv for 16, 48 or 90 hours and gene expression profile was obtained. There are in all 8 samples, two replcates of each 4. Two samples, namely 1D,1E (replicates of one) are control (PMA differentiated Thp-1 cells Uninfected controls). Cells were infected with H37Rv at an MOI of 1:10 and samples were collected at 16 hours, 48 hours and 90 hours post-infection.
Project description:Transcriptional profiling of Mtb H37Rv infected into THP-1 macrophage cell line and treated with 100 µM vitamin C (vit C) for 96 hours and 144 hours, compared to gene expression profile of untreated bacteria post-infection.
