Additional file 1 of Prohibitin 1 interacts with p53 in the regulation of mitochondrial dynamics and chemoresistance in gynecologic cancers
Bibliographic record
Abstract
Additional file 1 Figure S1. Protein-protein interaction analysis by PLA. Control of OVCA and CECA cell and description of Duo link PLA count method. (A) A2780s (ovarian cancer cells- OVCA), (B) C13 (cervical cancer cells – CECA), and (C) human ovarian tumour section without treatment of PLA reagents (contro. Blue represents DAPI (nucleus marker) and green represents TOM20 (mitochondria marker). (D) OV2008 cells treated without (CTL) or with CDDP were subjected to PLA assay. Using Duolink image tool, white dots (PLA signal) were counted either in mitochondria (green) or nucleus (Blue). Cell number was automatically assigned as shown by Duolink image tool. (E) PLA signal in each cell were counted, summed and averaged by number of cells as shown in table. Figure S2. CDDP increased Phb1 content and apoptosis in OV2008 cells, but not in C13* cells. (A) and (B) Comparison of Phb1 contents and apoptosis in OV2008 and C13* cultured with CDDP at different concentrations (A: 0–10 μM, 24 h) or for different duration (B: 0–24 h, 10 μM). Contents of Phb1 and GAPDH (loading control) were examined by Western blotting. Apoptosis was examined by Hoechst assay. Phb1 contents in OV2008 cells but not in C13* cells were significantly increased in the presence of CDDP in a concentration- (A; **p < 0.01 ***p < 0.001, n = 3) and time- (B; ***p < 0.001, n = 3) dependent manner. OV2008 cells exhibited higher apoptosis than C13* cells when treated with CDDP. Results are expressed as mean ± SEM (n = 3) and analyzed by 2-way ANOVA and Bonferroni post-hoc test. [**p < 0.01, ***p < 0.001, (versus CDDP = 0; A) and (versus time = 0; B); n = 3]. Figure S3. p-p53 (ser15) interacts with Phb1 and Bak in response to CDDP in chemosensitive CECA cells, but not in chemoresistant cells. (A) OV2008 and C13* cells were treated with CDDP (0–10 μM, 6 h). Protein contents of Phb1, p-p53 (ser15), p-p53 (ser20), Bak and GAPDH were examined by Western blot. Protein-protein interaction was determined by IP-Western. (B) Cell lysates were immunoprecipitated with IgG (control; lanes 1) or Bak antibody. Bak immunoprecipitates were immunoblotted [IP: anti-Bak, WB: anti-Bak, −Phb1, −p-p53 (ser15 and ser20)]. Results show representative images from 3 independent experiments. Results are expressed as mean ± SEM (n = 3) and analyzed by 2-way ANOVA and Bonferroni post-hoc test. [A and B; **p < 0.01, ***p < 0.001 (compared to DMSO), n = 3]. Figure S4. Prolonged CDDP treatment induced mitochondrial localization of Phb1 in CECA cells. (A) OV2008 and (B) C13* CECA cells were cultured with CDDP (0, 10 μM, 0, 3, 6, and 24 h; DMSO as a vehicle), and were examined by confocal microscopy. Cellular localization of Phb1 (Red) and p-p53 (ser15) were shown in representative images. Green: Mitochondrial Marker and Blue: Nucleus marker (DAPI) Merge 1 indicates the merged image between Phb1 and p-p53 (ser15) whereas merge 2 indicates merged image between DAPI and Phb1.
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How this classification was reachedexpand
Full frame machine prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. The Gemma side is a direct model label for every work in the frame, read from the title-only record. The Codex side is a classifier learned from the 10,348 direct Codex labels and calibrated to design-weighted sample rates; fields without enough sample support carry no Codex call. Candidate is the union of the two sides; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels.
Distilled classifier scores by category (both heads)
| Category | Codex | Gemma |
|---|---|---|
| Metaresearch | 0.002 | 0.020 |
| Meta-epidemiology (narrow) | 0.002 | 0.001 |
| Meta-epidemiology (broad) | 0.002 | 0.001 |
| Bibliometrics | 0.002 | 0.004 |
| Science and technology studies | 0.001 | 0.000 |
| Scholarly communication | 0.002 | 0.002 |
| Open science | 0.002 | 0.001 |
| Research integrity | 0.002 | 0.001 |
| Insufficient payload (model declined to judge) | 0.883 | 0.156 |
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 source (direct Gemma or distilled Codex), 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".