Argonaute 2 Silencing of Band 3 in Early Reticulocytes
Bibliographic record
Abstract
Abstract Post-transcriptional gene silencing by RNA-induced silencing complex (RISC) in erythropoiesis is a rapidly evolving field of study. Disruption of microRNA (miRNA) synthesis has been shown to hinder erythropoiesis and has been linked to disease. The presence of miRNA in circulating RBCs has recently been reported; however, clear understanding of their role and effectors has yet to be determined. In this study, freshly drawn RBC were density separated on discontinuous percoll gradients. The top two fractions, fraction 1 (F1) representative of the top layer containing the youngest, reticulocyte-like cells, and a second fraction with greater density and non-reticulocyte properties (F2), were isolated. Cell populations were analyzed to compare RBC protein levels using western blotting (WB). qRT-PCR was used to evaluate band 3 RNA. Argonaute (AGO2) RNA immunoprecipitation (RIP), was used to ascertain RISC mediated transcriptional repression of band 3 using qRT-PCR. Band 3 protein expression was elevated in the denser F2 compared to F1, in freshly drawn normal RBCs. RNA levels of band 3 complemented protein levels, as increased band 3 RNA was observed in F2. RIP experiments revealed elevated post-transcriptional control of band 3 through increased association of its RNA with AGO2 in F1 compared to F2 thereby, correlating with greatly reduced, sometimes undetectable protein levels of band 3 in F1. We confirm the presence of AGO2 in circulating RBCs. We report, for the first time, the post-transcriptional control of band 3, an integral RBC transmembrane protein, in circulating reticulocyte-like cells. These results suggest a possible subpopulation of reticulocytes, a pro-reticulocyte, with ongoing control of band 3 expression following enucleation, and a potential role of miRNA-RISC in band 3 pathology. Disclosures No relevant conflicts of interest to declare.
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How this classification was reachedexpand
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 | 0.000 | 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 itClassification
machine, unvalidatedMachine predicted; a candidate call from one teacher head, not a consensus.
How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".