Each well was coated with 2 ml of bottom agar mixture (media, 10% FBS, 1% agar).
The bottom agar layer stabbed by the test strains was subsequently overlaid with a thin layer of a soft agar containing the indicator strain.
When necessary, the bottom agar layer supporting the growth of the test strains was kept in chloroform vapors for 15 min and then overlaid with a soft agar containing the indicator strain.
In the cases when the bottom agar layer stabbed with the test strains was not treated with catalase, we have observed very extensive, often merging zones of M. catarrhalis strains growth inhibition.
Cells in log phase were mixed with 0.7% agar solution, and the mixture (3 mL) was poured onto 1.6% bottom agar plate containing 20 mL of BHI medium (Bacto™ Brain heart infusion 37 g/L, Cockeysville, MD, USA).
mitis Sm_11/5 2/5 7 Sm_13/39 3/5 5 4 6 Sm_18/56Sm_18/56 7/12 6 7 7 aIn the cases when the bottom agar layer stabbed with the test strains was not treated with catalase, we have observed very extensive, often merging zones of M. catarrhalis strains growth inhibition, or no growth of the indicator strain at all bSlash marks the repeat of experiments.
Petri dishes were poured using 25 ml 7H9 bottom agar (1.2% agar) per dish.
Bottom agar was prepared by adding desired treatments (e.g. 6.66 ng/ml TPA) to the 0.5% agar mix.
A six-well culture plate was coated with 2 ml bottom agar mixture (DMEM/F12 with 10% FBS, 0.6% agar).
Briefly, 104 cells were mixed with 0.3% soft agar and plated on top of 0.6% bottom agar seeded on each of a 60 mm plate.
Each well (35 mm) of a six-well culture dish was coated with 2 ml bottom agar mixture (DMEM, 10% (v/v) FCS, 0.6% (w/v) agar).
