Supplementary material for Sharp and Pogliano (1999) Proc. Natl. Acad. Sci. USA 96 (25), 14553-14558.
D. Scoring of SpoIIIE-GFP localization in fully engulfed sporangia
SpoIIIE-GFP localization | Strain genotype | |||
Class | Phenotype | Wild type | spoIIIE73-11 | spoIIIE36 |
i | Single septal focus | 7 (5%) | 44 (44%) | 75 (72%) |
ii | Single flanking focus | 22 (15%) | 3 (3%) | 2 (2%) |
iii | Single polar focus | 36 (25%) | 4 (4%) | 0 (0%) |
iv | Two foci: Septal/Flanking | 6 (4%) | 2 (2%) | 9 (9%) |
v | Two foci: Septal/Polar | 2 (1%) | 35 (35%) | 16 (15%) |
vi | Delocalized | 72 (50%) | 12 (12%) | 2 (2%) |
Total sporangia scored | 145 | 100 | 104 | |
Fig. 7.
Localization of SpoIIIE-GFP by immunofluorescence microscopy in wild-type (A), spoIIIE36 (B), and spoIIIE73-11 (C) at t3. (Left) Chromosomes stained with DAPI (blue) and SpoIIIE-GFP stained with anti-GFP (green). (Right) Membranes stained with FM 4-64 (red). In each field, arrows with roman numerals indicate the classes scored in D: i. Single fluorescent focus in the region of the polar septum. ii. Single fluorescent focus flanking the forespore. iii. Single fluorescent focus at the cell pole. iv. Two fluorescent foci, one at the polar septum and one flanking the forespore. v. Two fluorescent foci one at the polar septum and one at the cell pole. vi. Delocalized fluorescence, which we defined as more than two fluorescent foci or bright fluorescence surrounding the forespore. (White bars in A, B, and C are 2 mm long.)