Nh4pf6

IrCl₃ hydrate was purchased from Precious Metals Online. 1,2,3,4,5-Pentamethylcyclopentadiene, dibenzocyclooctyne-amine (DBCO-NH₂),2.0 M oxalyl chloride, triethylamine (TEA), and analytical grade sodium chloride were purchased from Sigma-Aldrich. 4,4′-Dimethyl-2,2′-bipyridine was obtained from Carbosynth, and NH₄PF₆ from Alfa Aesar. Solvents of laboratory grade were used for synthesis without any further purification. HPLC grade water and acetonitrile used for RP-HPLC experiments with the additive, HPLC grade trifluoroacetic acid (TFA) were obtained from Fisher Scientific. Roswell Park Memorial Institute (RPMI-1640) medium, with 10% fetal bovine serum (FBS), and 1% v/v penicillin/streptomycin was used in the cellular experiments. The cyclic azide-functionalized peptide Azhx-c(CRWYDENAC) was purchased from Cambridge Research Biochemicals (Billingham, UK) with a purity of > 95% (HPLC/MS) with all L-configured amino acids in the sequence.

All solvents were of analytical grade (>95% purity). All buffer components were of biological grade and used without further purification. Iridium chloride hydrate, ppy, dfppy, H₂dcbpy and NH₄PF₆ were purchased from Alfa Aesar. Cisplatin, rapamycin, DMSO, PLE, PI, NAC, MTT, H₂DCFDA, JC-1, 3-MA, CCCP, CQ, AO and Annexin V-FITC apoptosis detection kit were purchased from Sigma-Aldrich. MTDR was purchased from Invitrogen. Lipofectamine^® 3000 was purchased from Thermo Fisher Scientific. Hoechst 33342 and z-VAD-fmk were purchased from Beyotime. Caspase-Glo^® 3/7 assay kit and CellTiter-Glo^® luminescence cell viability assay kit were purchased from Promega.
Ligands L₁ (dimethyl 2,2′-bipyridine-4,4′-dicarboxylate), L₂ (diethyl 2,2′-bipyridine-4,4′-dicarboxylate), L₃ (dibutyl 2,2′-bipyridine-4,4′-dicarboxylate) and L₄ (diisobutyl 2,2′-bipyridine-4,4′-dicarboxylate) were synthesized according to reported procedures24 . The cyclometalated Ir(III) chloro-bridged dimers [Ir(ppy)₂Cl]₂ and [Ir(dfppy)₂Cl]₂ were prepared according to literature methods57 58 (link). The purity of synthesized complexes was analysed via combustion analysis and was found to be ≥ 95% pure. All the tested complexes were dissolved in DMSO just before the experiments, and the final concentration of DMSO was 1% (v/v).

RuCl₃·nH₂O (J&K), bpy (J&K), phen (J&K), DIP (J&K), Re(CO)₅Cl (Sigma Aldrich), silver trifluoromethanesulfonate (Alfa Aesar), 4-pyridinecarboxaldehyde (Alfa Aesar), 5,6-diamino-1,10-phenanthroline (Alfa Aesar), NH₄PF₆ (Alfa Aesar), 4% paraformaldehyde (Beyotime), crystal violet (Beyotime), Dulbecco’s Modified Eagle Medium (DMEM, Gibco), Roswell Park Memorial Institute 1640 (RPMI 1640, Gibco), Fetal bovine serum (FBS, Gibco), penicillin-streptomycin (Gibco), MTT (J&K), Rh123 (J&K), H₂DCFDA (J&K), Hoechst 33342 (J&K), Annexin V-FITC Apoptosis Detection Kit (Beyotime). Primary antibodies against caspase-3 and PARP were purchased from Cell Signaling Technology. RuRe-1, RuRe-2 were dissolved in DMSO just before the experiments, and the concentration of DMSO in biological experiments was 1% (v/v). Cisplatin was dissolved in 0.9% sodium chloride solution just before use.
A LCQ DECA XP spectrometer was used for obtaining ESI-MS spectra. A Bruker Avance 600 spectrometer was used for obtaining ¹H NMR spectra. A SpetraMax M2 plate reader was used for determining cell viability. A Nikon A1R/A1 laser-scanning confocal microscope was used for obtaining cell imaging images. A CyFlow Space flow cytometer was used for performing the flow cytometry analysis.