Whatman filter 1

Cholesterol assimilation and dry weight experiments for each growth condition were prepared in triplicate, where three independent sets of culture were homogenized and divided into six sterile 50 mL borosilicate tubes to account for six timepoints (0, 24, 36, 48, 60, and 72 h). At designated time point, a 1.0 mL aliquot of culture supernatant was collected, centrifuged at 2000× g for 15 min, and stored in a 50 mL borosilicate glass tube with a PTFE-lined cap at −20 °C. The remaining 9.0 mL of the culture was then harvested and filtered via vacuum flask and a pre-weighed Whatman filter #1. The contents were allowed to air dry for five days and dry weight was measured on an analytical balance.

Cholesterol assimilation and dry weight experiments were prepared in duplicate, where two independent sets of cultures were homogenized and divided into three sterile 50 mL borosilicate tubes to account for three timepoints (0, 24, and 72 h). M. purpureus was incubated in PBS buffer supplemented with either ammonium sulfate or yeast nitrogen base without amino acids, pH 7 with 0.3% oxgall and 120 µg/mL cholesterol. At designated time points, a 1.0 mL aliquot of culture supernatant was collected, centrifuged at 2000× g for 15 min, and stored in a 50 mL borosilicate glass tube with PTFE lined cap at −20 °C. M. purpureus samples were then washed twice under sterile conditions with 10 mL of PBS + 0.3% oxgall, and transferred to 10 mL MEA media with 0.3% oxgall. At day 4, the culture was harvested and filtered via vacuum flask and a pre-weighed Whatman filter #1. The contents were allowed to air dry for five days and dry weight was measured on an analytical balance.

Cholesterol assimilation and dry weight experiments were prepared in triplicate, where three independent sets of cultures were homogenized and divided into five sterile 50 mL borosilicate tubes to account for five timepoints (0, 24, 48, 72, and 96 h). M. purpureus was incubated in PBS buffer, pH 7 with 0.3% oxgall and 120 µg/mL cholesterol. At designated time points, a 1.0 mL aliquot of culture supernatant was collected, centrifuged at 2000× g for 15 min, and stored in a 50 mL borosilicate glass tube with a PTFE-lined cap at −20 °C. M. purpureus samples were then washed twice under sterile conditions with 10 mL of PBS + 0.3% oxgall, and transferred to 10 mL MEA media with 0.3% oxgall and 120 µg/mL cholesterol. A 1.0 mL aliquot of culture supernatant was collected at the starting point (t = 0 h), and also at day 4 and day 7 of incubation in MEA media. At day 7, the culture was harvested and filtered via vacuum flask and a pre-weighed Whatman filter #1. The contents were allowed to air dry for five days and dry weight was measured on an analytical balance.