Extended Data Fig. 9: Experimental data supporting the validity of the prodrug-based combination therapy, mediated by compartmentalized HS–E. coli whole-cell catalysts.

a, An illustration showing the setup of the experiment, in which Cu–HS–E. coli and Pd–HS–E. coli-mediated CuAAC and Proc-removal reactions in live mice. b, The images of live HS–E. coli containing fluorescent products in the mouse feces, collected during the experiment described in a. These results clearly demonstrated the activity of such whole-cell catalysts in mouse intestines. See the Whole-cell catalysis by compartmentalized, ArM-containing HS–E. coli in mouse intestine subsection in the Supplementary Information for detailed protocol. n = 5 biological replicates for each reaction; representative results are shown. Scale bar = 5 µm. c, Heatmap showing the cytotoxicity of doxorubicin (Dox, IC₅₀ = 4 µM), farnesyl umbelliferone (FUmb, IC₅₀ = 64 µM) and their combinations toward HCT116 cells. The experiment found Dox and FUmb showing the highest synergy at a 1:32 (wt/wt) ratio. At this ratio, the IC₅₀ of Dox/FUmb combinations was measured to be 0.7 µM (Dox) + 23 µM (FUmb) through curve-fitting. d, Schematic illustration on the in vitro validation of Pd and Ru whole-cell catalyst’s capability of activating Pro-Dox and Pro-FUmb. Prodrugs were added to solution containing HS–E. coli catalysts, and after 24 h, the supernatant was collected for HPLC characterization. The prodrug activation reactions are shown on the right. e, HPLC analysis results indicated that the whole-cell catalysts successfully mediated the desired abiotic transformation in the experiment depicted in d, and the products found in the supernatant indicated that the activated drugs could diffuse out of the bacterial cells. Concentrations used during incubations: 25 µM for Pd, 100 µM for Ru, 10 µM for Pro-Dox, 100 µM for Pro-FUmb, whole-cell catalyst OD = 0.6.