We examined (i) whether the adhesion of UPEC to their urothelial receptors in vivo is a prerequisite for altering the activation status of NF-κB, (ii) whether distinct urothelial layers respond differently to T1F-UPEC, (iii) whether the multiple nuclei within a single umbrella cell react synchronously or non-synchronously to T1F-UPEC infection, (iv) whether NF-κB activation is dependent on Toll-like receptor 4 signaling and (v) whether inhibiting NF-κB affects the outcome of urinary tract infections. Here, NFKB1 is linked to urinary tract infection.