DEV032607 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1. SDF-3 purification on HPLC. About 22,000 units of partially purified SDF-3 were loaded on a Majic C-18 column using an ultrafast HPLC apparatus and eluted with a methanol gradient. The LC mobile phase A was 3% methanol in water and the LC mobile phase B was 100% methanol. The LC flow rate was 50 µl/minute, and the LC gradient was 90% A to 95% B for 20 minutes, then held at 95% B for 5 minutes. The theoretical methanol gradient, calculated with a void volume of 125 µl, is indicated in red. Fractions of 50 µl were collected and tested by bioassay. Only fractions at 20 and 21 minutes had significant activity. The SDF-3 total activity in each fraction was calculated after serial dilution and is shown with the fraction (gray columns). The elution times for nine steroids were determined in a separate run on the same column using identical elution conditions. Peaks were detected at 240 nm (UV) and the steroids identified using a mass spectrometer coupled to the HPLC column. 1, aldosterone; 2, hydrocortisone; 3, dexamethasone; 4, 11-deoxycortisol; 5, corticosterone; 6, 11α progesterone; 7, deoxycorticosterone; 8, 17α progesterone; 9, progesterone.

• Supplemental Figure S2 -

  Fig. S2. Sporulation induced by steroids. Developed KP cells were incubated with various concentrations of the indicated steroids. The number of spores was determined after 1 hour of incubation. Each experiment was repeated three to five times. The results are split into two plots (A,B) for clarity.

• Supplemental Table S1 - Adobe PDF

• Supplemental Table S2 - Adobe PDF