| Category |
Data Type |
Description |
Notes |
| No Clones |
Yes/No |
True (-1), False (0) |
When ‘TRUE’, the animal presents no GFP+ clones and therefore can not be described in any of the other categories. If 5 consecutive animals show no clones, then no more animals are screened. |
| Not Developed |
Yes/No |
True (-1), False (0), True if pupa was too malformed to mount |
When ‘TRUE’, the animal has not developed sufficiently within the normal time window and therefore can not be described in any of the other categories. If 5 consecutive animals show a developmental delay, then no more animals are screened. |
| Malformed |
Yes/No |
Normal (0), malformed (1) |
When ‘Malformed’, the notum appeared malformed under the stereomicroscope and was sometimes difficult to mount. |
| Freckles |
Yes/No |
No (0), Yes (1) |
When ‘Yes’, one or more necrotic dark spots were visible in the notum under the stereomicroscope. |
| Clonal tissue |
Number |
0-5% (-2), 5-15% (-1), 15-35% (0), 35-50% (1), 50% + (2) |
‘Clonal tissue’ refers to the proportion (%) of notum that is positively labelled with GFP. In a wild-type notum, it is typically 50%. |
| Individual clone size |
Number |
mostly small (-1), mixture/mostly medium (0), mostly large (1), very large (2) |
‘Individual Clone Size’ refers to the size of individual clones in the notum. |
| Clonal shape |
Number |
mostly round (-1), mixture (0), mostly long (1) |
‘Clonal shape’ referes to the shape of individual clones in the notum. |
| Dividing cells |
Number |
none (-2), little (-1), medium (0), lots (1), extensive (2) |
‘Dividing cells’ refers to the number of dividing cells in the notum. We considered dividing cells all cells with a rounded and enlarged morphology just prior to cell division, and daughter cells as they undergo cytokinesis. |
| Invading Cells |
Number |
none (-2), very few (-1), medium (0), lots (1), extensive (2) |
‘Invading cells’ refers to the number of cells that have delaminated from the epithelium. |
| Type of Invasion |
Number |
mostly non-polarised (-1), mixture (0), mostly polarised (1) |
‘Type of invasion’ refers to the morphology of cells that have delaminated, ie. ‘polarised’ are round cells with a GFP enriched spot on the cortex, ‘non-polarised’ are round cells without a GFP spot. |
| Invading cells with protrusions |
Yes/No |
No (0), Yes (1) |
When ‘Yes’, ‘Invading cells with protrusions’ are seen in the notum. These are basally delaminated cells that present membrane protrusions, either or both lamellipodia and filopodia and often present a polarised movement on X/Y or Z planes. |
| Cuticle |
Yes/No |
normal (0), deep folding (1) |
The cuticle is an autofluorescent structure secreted apically by the epithelial cells. If deep folding is present in the cuticle, it can be suggestive of epithelial folding, which typically occurs when there is overproliferation of the epithelial sheet. |
| closure defects |
Yes/No |
No (0), Yes (1) |
When ‘Yes’, it indicates that the two (left and right) developing epithelial sheets have not fully joined at the midline. |
| Apex size |
Number |
very small (-2), small (-1), normal (0), large (1) |
‘Apex size’ refers to the size of the apex in the majority of the GFP+ cells. |
| Defective Apex |
Number |
none (0), up to 50% (1), >50% (2) |
‘Defective apex’ refers to the proportion of GFP+ cells that presented an atypical apex morphology. |
| No Apex |
Number |
none (-1), 0-15% (0), 15-50% (1), >50% (2) |
‘No Apex’ refers to the proportion of GFP+ cells that presented no apex, often as a consequence of losing their apicobasal polarity. |
| Junction defects |
Yes/No |
No (0), Yes (1) |
When ‘Yes’, ‘Junction defects’ flags the absence of defined junctions or the presence of morphologically defective junctions. |
| Cell body (rounding) |
Number |
none (-1), 0-15% (0), 15-50% (1), 50% (2) |
‘Cell body rounding’ refers to the proportion of cells within the epithelial sheet that present a loss of apicobasal polarity and appear with a rounded morphology. |
| Polarised Cytoskeleton |
Number |
none (-1), very few (0), medium (1), lots (2) |
‘Polarised cytoskeleton’ refers to the numbers of cells within the epithelial sheet that appear with a rounded morphology and a GFP+ enriched spot on the cortex. These are typically cells that are about to delaminate basally. |
|
Yes/No |
No (0), Yes (1) |
When ‘Yes’, ‘Basal bundles’ refers to very long membrane protrusions at the base of epithelial cells that present in tight and thick bundles. |
| Long apical/intermediate protrusions |
Yes/No |
No (0), Yes (1) |
When ‘Yes’, ‘Long apical/intermediate protrusions’ refers to the presence of long membrane protrusions that resemble filopodia at the apex or in middle sections of epithelial cells. In the wild-type animal, long protrusions are typically present only at the base of the cell. |
| Very Long protrusions |
Yes/No |
No (0), Yes (1) |
When ‘Yes’, ‘Very long protrusions’ indicates the presence of unusually long membrane protrusions at the base of epithelial cells. |
| Blebbing mitotic cells |
Yes/No |
No (0), Yes (1) |
When ‘Yes’, ‘Blebbing mitotic cells’ refers to the presence of cells undergoing cell division that present membrane blebbing. |
| Thick protrusions |
Yes/No |
No (0), Yes (1) |
When ‘Yes, ‘Thick protrusions’ refers to the presence of thick basal protrusions. |
| Bright protrusions |
Yes/No |
No (0), Yes (1) |
When ‘Yes, ‘Bright protrusions’ referes to the presence of unusually bright GFP+ basal protrusions. This may indicate actin enrichment within these protrusions. |
| Branched protrusions |
Yes/No |
No (0), Yes (1) |
When ‘Yes, ‘Branched protrusions’ refers to the presence of unusually branched basal protrusions. |
| Cyst-like clones |
Yes/No |
No (0), Yes (1) |
When ‘Yes’, Cyst-like clones are observed. These are hollow spherical structures, where an epithelial sheet folds in on itself to form a ball with a lumen. |
| Apoptotic |
Number |
None (0), little (1), medium (2), lots (3) |
‘Apoptotic’ refers to the numbers of epithelial cells that appeared to be undergoing apoptosis. This was assumed based on abnormal cell morphology together with a loss of GFP intensity. |
| Basal Protrusions |
Number |
absent (-3), small (-2), normal (-1), long (0), very long (1) |
‘Basal protrusions’ refers to the absence or length of membrane protrusions at the base of epithelial cells. |
| Basal Actin Rich Spot |
Number |
No (0), Yes (1) |
‘When ‘Yes’, ‘Basal actin rich spot’ refers to the presence of GFP accumulation in a spot within the basal protrusions. |
| Multi-layering |
Number |
none (-1), 0-15% (0), 15-50% (1), 50+% (2) |
‘Multilayering’ refers to a mutilayered epithelium as opposed to the wild-type monolayered epithelium. |
| Cell Length |
Number |
short <7μm (-2), normal 8-12μm (-1), long (0) 13-19μm, very long (1) 20μm + |
‘Cell length’ refers to the length of epithelial cells in the z (apicobasal) axis |
| Single Cell Clones |
Yes/No |
No (0), Yes (1) |
When ‘Yes’, ‘Single cell clones’ refers to the presence of clones of only one cell. |
