Down-regulation of podoplanin expression in TPC1 cells following transfection with PDPN-specific siRNA.
Bibliographic record
Abstract
<p>A. RT-qPCR analysis of PDPN mRNA levels in TPC1 thyroid cancer cells 48 h after transfection with 30 nM siRNA specific for PDPN (siPDPN) or a negative control siRNA (siNEG). The results were normalized to the 18S rRNA level and bars represent the average fold change in PDPN transcript abundance in cells transfected with siPDPN compared with cells transfected with siNEG. The results are representative of four independent experiments. Data are presented as the mean ±SEM, **<i>P</i><0.001. B. Western blot analysis of podoplanin and β-actin proteins in TPC1 cells before (0 h) and 48 h after transfection with siPDPN and control siNEG. C. Immunofluorescence staining of podoplanin protein in TPC1 cells transfected with siPDPN and control siNEG. Cells were stained with anti-PDPN monoclonal antibody D2-40 followed by DyLight549-conjugated secondary antibody (red), and counterstained with DAPI (blue). Magnification 1000x. D. Effect of PDPN on cell viability. D, left panel. Proliferation was measured at 24 and 48 h after transfection of TPC1 cells with siPDPN or control siNEG. Cells seeded in 96-well plates were treated with XTT mixture reagent and formazan formation was measured at 450 nm to determine the number of viable cells. Data are expressed as the mean ±SEM of at least three independent experiments performed in quintuplicate. D, right panel. Apoptosis was measured at 48 h after transfection of TPC1 cells with siPDPN or control siNEG. Cells were collected and stained with FITC Annexin V and propidium iodide, followed by flow cytometry. Representative measurements of the percentage of Annexin V+ cells are presented. Each bar represents the mean ±SEM of at least three independent experiments performed in quadruplicate. E. Function of podoplanin in cell adhesion. TPC1 cells transfected with siPDPN display adhesion capacity comparable to those of siNEG-transfected cells. Briefly, 8000 transfected cells were seeded in the wells of 96-well plates. After incubation for 24 or 48 h, the cell monolayers were washed, fixed with 4% formaldehyde for 15 min and stained with crystal violet (Merck, USA). The stained cells were lysed by treatment with 2% SDS, then the intensity of the released stain was quantified by spectrophotometry at 550 nm using a Labsystems Multiscan RC microplate reader (Thermo Fisher Scientific, Canada). Data represents three separate experiments.</p>
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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) | 0.009 | 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".