Nature Microbiology 2025 (Jul); 10 (7): 1630–1647

Extended Data Fig. 3. PFAS bioaccumulation in cell pellet.

A. Data from Fig. 2c including compound control and whole culture samples. Accumulation of PFDA and PFNA by wild-type E. coli strains and corresponding efflux mutants. OD₆₀₀ = 3.75; exposure concentration = 20 µM; two-sided t-test; ** p-value < 0.01 and >20% sequestration compared to the compound control; *** p-value < 0.001 and >20% sequestration compared to the compound control; n = 3 technical replicates (Supplementary Table 19).

B. Efflux mutants show increased PFAS sensitivity. n = 3 technical replicates (Supplementary Tables 16, 17).

C. Independent measurement of samples from Fig. 2d at Imperial College London supports PFNA accumulation by B. uniformis at 160 ng/l exposure concentration. Two-sided t-test (supernatant compared to the compound control; pellet compared to pellet control); * p-value < 0.05; ** p-value < 0.01; n = 3 biological replicates (Supplementary Table 34).

D. Data from Fig. 2d displayed in a different way: based on wet pellet weight, concentration within the bacterial pellet was a median of circa 8200 ng/l, which is a 50-fold increase compared to the exposure concentration. Two-sided t-test (supernatant compared to the compound control; pellet compared to pellet control); * p-value < 0.05; ** p-value < 0.01; n = 3 biological replicates (Supplementary Tables 20, 21).

E. Data related to Fig. 2e: PFAS recovery from pellet and supernatant after exposure to 5 µM PFAS in mGAM (initial OD₆₀₀ = 0.05, 24 h incubation) or PBS (OD₆₀₀ = 3.75, 4 h incubation). Bars depict median percent recovery from the bacterial pellet, supernatant or compound control; error bars depict standard error; overlaid points depict single data points; n = 3 technical replicates; (Supplementary Table 35).

F. Data from Fig. 2f including compound control, whole culture and supernatant samples. PFAS recovery after 1 h exposure in PBS (OD₆₀₀ = 3.75, PFAS mix of 14 compounds each at a concentration of 1 mg/l). For PFAS compounds with a chain length of more than ten carbon atoms, the solubility is limited at this concentration; the low accumulation in E. coli BW25113 wild-type pellets acts as a biological control in this case. Bars depict median percent recovery from the compound control, whole culture, supernatant or bacterial pellet; error bars depict standard error; overlaid points depict single data points; n = 3 technical replicates; (Supplementary Table 36).