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. 2023 Feb 16;24(4):e56325. doi: 10.15252/embr.202256325

Figure EV1. Wild‐type p53 activates METTL14 expression in CRC cell lines.

Figure EV1

  1. METTL14 mRNA levels between tumor and normal tissues in the TCGA database (*P < 0.05, **P < 0.01, ***P < 0.001). Breast cancer (BRCA—normal tissues n = 113, cancer tissues n = 1,104); colon cancer (COAD—normal tissues n = 41, cancer tissues n = 471); rectal cancer (READ—normal tissues n = 10, cancer tissues n = 167); endometrioid cancer (UCEC—normal tissues n = 35, cancer tissues n = 548); kidney clear cell carcinoma (KIRC—normal tissues n = 72, cancer tissues n = 535); lung adenocarcinoma (LUAD—normal tissues n = 59, cancer tissues n = 526); lung squamous cell carcinoma (LUSC—normal tissues n = 49, cancer tissues n = 501); prostate cancer (PRAD—normal tissues n = 52, cancer tissues n = 499); and thyroid cancer (THCA—normal tissues n = 58, cancer tissues n = 510).
  2. qRT–PCR determination of the METTL14 level in CRC and adjacent normal tissues from Cohort 1 (n = 118).
  3. Representative IHC staining images and statistical analysis of METTL14 in tumor and para‐tumor tissues from Cohort 2 (n = 90). Lower panels show enlarged images of indicated normal or CRC tissues. Scale bars = 200 μm (upper) and 40 μm (lower).
  4. qRT–PCR analysis of the mRNA levels of METTL14 after transfection with control or ELK1 siRNAs with 48 h treatment in HCT116 and Lovo cells. Data are presented as mean ± SD (biological replicates, n = 3; *P  < 0.05).
  5. Western blot analysis of METTL14 and ELK1 protein levels after transfection with control or ELK1 siRNAs with 48 h treatment in HCT116 and Lovo cells.
  6. Expression analysis of METTL14 in p53‐WT and p53‐MT or KRAS‐WT and KRAS‐MT tumor tissues from in TCGA. Lung Cancer (LC); Head and Neck Cancer (HNSC); Stomach Cancer (STAD); Liver Cancer (LIHC) and Pancreatic Cancer (PAAD).
  7. HT29 (p53R273H), SW620 (p53R273H), COLO320 (p53R248W) and SW480 (p53R273H/P309S) cells were transfected with control or p53 siRNAs. Forty‐eight hours after transfection, total protein was then analyzed by western blot analysis. Arrows indicate METTL14 protein.
  8. Representative IHC images, and statistical analysis of immunoreactive score (IRS) of METTL3 expression in p53‐WT (n = 63) and p53‐MT (n = 41) CRC samples from Cohort 3. The insets show enlarged images of indicated p53‐WT and p53‐MT CRC tissues, respectively. Scale bars = 20 μm and 2 μm (inset). The horizontal lines represent the median; the bottom and top of the boxes represent the 25 and 75% percentiles, respectively, and the vertical bars represent the range of the data.
  9. Schematic illustration of putative p53‐binding site in METTL14 gene promoter.

Data information: For (A, B, D, and F), statistical significance was determined by a two‐tailed Student's t‐test. For (H), statistical significance was determined by the nonparametric Mann–Whitney test. For (C), statistical significance was determined by the χ2 test. ACTB was used as a loading control.