Monitoring Autophagy in <i>Drosophila</i> Using Fluorescent Reporters in the UAS-GAL4 System
Why this work is in the frame
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Bibliographic record
Abstract
Following autophagy induction, the autophagy-related protein Atg8 undergoes ubiquitin-like conjugation to phosphatidylethanolamine and inserts into the autophagosome membrane. Transgenic Drosophila lines expressing Drosophila Atg8 (DrAtg8a) fused to green fluorescent protein (GFP), mCherry, or dual-tagged GFP-mCherry have been constructed and are extremely useful for monitoring autophagy. The use of GFP-mCherry-Atg8a is particularly advantageous because it allows for the assessment of autophagy induction as well as autophagy flux. GFP-mCherry-Atg8a fluoresces yellow in nonacidic structures including the autophagosome. When autophagosomes fuse with lysosomes to form autolysosomes, GFP fluorescence is quenched by the acidic hydrolases and the resulting autolysosome will fluoresce red. The upstream activating sequence (UAS)-GAL4 system allows for the ectopic expression of the gene of interest (in this case, DrAtg8a) in a tissue-specific manner. Here we provide a protocol for monitoring autophagic flux by fluorescence microscopy by expressing UASp-GFP-mCherry-DrAtg8a in the Drosophila germline.
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Full frame distilled prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. Learned from the 10,348 direct Codex labels and 10,348 direct Gemma labels. Candidate is the union of thresholded teacher heads; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels or direct frontier model labels.
Codex and Gemma teacher scores by category
| Category | Codex | Gemma |
|---|---|---|
| Metaresearch | 0.001 | 0.000 |
| Meta-epidemiology (narrow) | 0.000 | 0.000 |
| Meta-epidemiology (broad) | 0.000 | 0.000 |
| Bibliometrics | 0.000 | 0.000 |
| Science and technology studies | 0.000 | 0.000 |
| Scholarly communication | 0.000 | 0.000 |
| Open science | 0.000 | 0.000 |
| Research integrity | 0.000 | 0.000 |
| Insufficient payload (model declined to judge) | 0.000 | 0.000 |
Machine scores (provisional)
The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.
Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.
score_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from it