Fig 1: LATS2 expression is down-regulated by miR-15b-5p during prostate cancer development. (A) Western blot was performed to detect LATS2 protein expression in prostate cancer tissues. (B) LATS2 was down-regulated in prostate cancer PC-3 cells. (C) MiR-15b-5b restricted the expression of LATS2. (D) The expression of LATS2 was up-regulated following inhibition of miR-15b-15b. Comparisons were performed using the paired t-test. *Indicates p <0.05, **Indicates p <0.01. Error bars represent SEM. Data represent three independent experiments.
Fig 2: Silence of LATS promotes miR-15b-5b-induced cell proliferation. (A) The expression of LATS2 protein in PC-3 cells with or without LATS2 suppression following miR-15b-5b overexpressing was detected by Western blot. (B) Cell Counting Kit-8 was used to evaluate the proliferation of PC-3 cells with or without LATS2 suppression following miR-15b-5b overexpressing. (C) Colony formation assay was performed to evaluate the colony-forming capacity of PC-3 cells with or without LATS2 suppression after miR-15b-5b overexpression (Magnification:1×); (D) LATS suppression promoted PC-3 cells proliferation induced by miR-15b-5b. Comparisons were performed using the one-way ANOVA. *Indicates p <0.05, **Indicates p <0.01. ***Indicates p <0.001. ****Indicates p <0.0001. Error bars represent SEM. Data represent three independent experiments.
Fig 3: Validation of the differentially expressed genes by reverse transcription-quantitative PCR and western blot. (A) mRNA levels of Lats2, Lats1 and Tnfrsf11b. (B) Protein levels of Lats2, Lats1 and Tnfrsf11b. Lanes 1–3 are for individual mice from the S02 group and lanes 4–6 from the S01 group. *P<0.05 vs. S01. Groups: S01, Control group; S02, Infected group. Lats, large tumor suppressor kinase; Tnfrsf11b, TNF receptor superfamily member 11b.
Fig 4: Overexpression of LATS2 attenuates miR-15b-5b-induced cells proliferation. (A) Western blot was performed to detect the expression of LATS2 protein in PC-3 cells with or without LATS2 overexpression following overexpression of miR-15b-5b. (B) The proliferation of PC-3 cells transfecting with the indicated vectors was analyzed by CCK8. (C) The colony-forming capacity of prostate cancer PC-3 cells with or without LATS2 overexpression after miR-15b-5b overexpression was evaluated by colony formation assay (Magnification:1×). (D) The proliferation of PC-3 cells induced by miR-15b-5b was inhibited by LATS2. Comparisons were performed using the one-way ANOVA. *Indicates p <0.05, **Indicates p <0.01. ***Indicates p <0.001. ****Indicates p <0.0001. Error bars represent SEM. Data represent three independent experiments.
Fig 5: LATS2 acts as a target of miR-15b-5b and negatively regulated by miR-15b-5p. (A) miRanda was used to predict miR-15b-5b targeted to WT or Mut 3'-UTR of LATS2. (B) After miR-15b-5b overexpression, luciferase activity assay was performed to detect the luciferase activity of reporters containing WT or Mut 3'-UTR of LATS2 with miR-15b-5p target sites in RWPE-1 cells. (C) The same method was used to detect the luciferase activity in prostate cancer PC-3 cells. Comparisons were performed using the paired t-test. ****p <0.0001. NS indicates no significant difference. Error bars represent SEM. Data represent three independent experiments.
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