human bladder epithelial cell atcc htb  (ATCC)

Journal: bioRxiv

Article Title: Lysine Decarboxylation aids in UPEC intracellular survival in the early stages of urinary tract infection

doi: 10.1101/2025.04.25.650641

Figure Lengend Snippet: Graph shows RT-qPCR results of representative genes from each AR system, using RNA isolated from intracellular bacteria. For this work the bladder epithelial cell line 5637 (ATCC HTB-9) was seeded with wild-type UTI89 at an MOI=10. Following a 2h incubation to allow for pathogen internalization, extracellular bacteria were eliminated by incubating the urothelial cell line with gentamicin for another 2 hours. Gentamicin was then removed and cells were washed 3X with 1X PBS, followed by urothelial cell lysis and subsequent treatment for bacterial lysis and RNA extraction. Extracted RNA was DNAse-treated, reverse-transcribed, quantified and subjected to qPCR using TaqMan probes designed for adiA (purple), cadA (green), gadA (pale blue), gadC (light blue), sdaA (red), sdaC (royal blue), and speF (coral). Transcript abundance was normalized to the gyrB house keeping gene transcripts. Relative fold changes were determined by the ΔΔ C T method, relative to the bacterial inoculum corresponding transcripts. Graph shows analysis of 4 biological replicates. Error bars indicate mean and standard error of the mean at 95% CI.

Article Snippet: Cell culture infections were performed using 5637 human transitional bladder epithelial cells (ATCC HTB-9) originally derived from a 68-year old white male with Grade II bladder carcinoma.

Techniques: Quantitative RT-PCR, Isolation, Bacteria, Incubation, Lysis, RNA Extraction, Reverse Transcription