Bladder cells are constantly exposed to multiple chemical stressors stemming from urine, including xenobiotics and bioactive metabolites. Even if not completely understood at the molecular level, a significant adaptability to these challenges is a prerequisite for bladder cells and enables the maintenance of physiological function. Here untargeted profiling was applied to grasp changes in bladder cells proteome upon application of tailored chemical interventions. To focus on regulatory events observable in the nuclear and peri-nuclear compartments, nuclear extracts were generated after incubation with different molecules enabling regulation of oxidative stress (1 µM MitoTEMPOL, 500 µM H2O2), autophagy (10 nM Bafilomycin A1), cytoskeletal integrity (100 µM CK666), and metabolism (1 mM 2-desoxyglucose). T24 cells were incubated with the respective treatments for 24 hours, after which nuclear proteins were isolated, digested and analyzed using a timsTOF Pro mass spectrometer (Bruker). Identification as well as label-free quantification (LFQ) of proteins and statistical analyses were performed using publicly available software package MaxQuant 1.6.17.0.