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. Author manuscript; available in PMC: 2023 Jun 20.
Published in final edited form as: Nat Cell Biol. 2023 Jan 16;25(2):309–322. doi: 10.1038/s41556-022-01055-y

Fig. 3: Stress suppresses miR-139-5p to release the brake on LPAR4 expression.

Fig. 3:

a, Overall survival probability for high vs. low miR-139-5p for the TCGA PAAD dataset. b, Eight oligonucleotides are paired between LPAR4 3’UTR and hsa-miR-139-5p. c, Relative mRNA levels of LPAR4, miR-139-5p, and miR-138-5p in Colo-357 cells treated with serum deprivation stress (n= 3 independent experiments), hypoxia (n=3 independent experiments except n=2 for miR-138-5p), or oxidative stress (n= 4 independent experiments), normalized to cells treated with no stress. d, Relative mRNA levels of miR-139-5p and miR-138-5p in secondary methylcellulose spheres grown from Colo-357 or 34E cells as compared to their expressions in cells grown on 2D. e, Relative mRNA level of LPAR4 in Colo-357 (n=7 independent experiments) or 79E cell lines (n=4 independent experiments) treated with scrambled control miRNA or miR-139-5p mimic in normoxia vs. hypoxia. f, LPAR4 protein level in three cell lines treated with scrambled control miRNA or miR-139-5p mimic in normoxia vs. hypoxia. Data are representative of three independent experiments. g, The mRNA level of LPARs and FOXO1 in 34E cells treated with miR-139-5p inhibitor relative to cells treated with scrambled control (n=3 independent experiments). h, The relative luciferase values in 34E (n=3 independent experiments) or Colo-357 (n=4 independent experiments) treated with miR-139-5p mimic or anti-miR-139-5p, along with their scrambled controls. i, Impact of miR-139-5p inhibitor on the mRNA level of LPAR4 and the number of tumorspheres in Colo-357 cells with or without LPAR4 knockdown (n=3 independent experiments for tumorsphere assay and n=4 for quantitative RT-PCR assay). j, Impact of miR-139-5p mimic on the mRNA level of LPAR4 and on the number of tumorspheres in Colo-357 cells with or without LPAR4 ectopic expression (n=3 independent experiments for tumorsphere assay and n=5 for quantitative RT-PCR assay). Data are presented as mean ± s.d. in c-e and g-i. Statistical analyses were performed using two tailed unpaired one sample t-test (c-e and g-j). Source numerical data and unprocessed blots are available in source data.