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. 2021 Oct 14;22(12):e52124. doi: 10.15252/embr.202052124

Figure 6. LncRNA CRNDE negatively regulated PTEN expression.

Figure 6

  • A, B
    (A) RNA Immunoprecipitation (RIP) and (B) RNA‐pull down assays were conducted to detect the association of LncRNA CRNDE and phosphatase and tensin homolog (PTEN)/neural precursor cell expressed developmentally downregulated protein 4‐1 (NEDD4‐1).
  • C
    SGC7901 cells were transfected with pcDNA‐CRNDE or its negative control (Mock) before MG132 treatment (10 µM, 6 h). The co‐immunoprecipitation (Co‐IP) assay was performed to examine the combination of PTEN and NEDD4‐1.
  • D
    The expressions of PTEN and NEDD4‐1 were measured using Western blot in SGC7901 cells transfected with pcDNA‐CRNDE or Mock. GAPDH was served as an internal control.
  • E
    The expressions of PTEN and NEDD4‐1 were measured in SGC7901 cells transfected with pcDNA‐CRNDE or Mock at four time‐points after cycloheximide (CHX, 100 μg/ml) treatment. GAPDH was served as an internal control. Left: Representative bands. Right: Quantitative results. The results shown are from three biological replicates and are expressed as mean ± SD. Student’s t‐test. *P < 0.05, **P < 0.01 vs. Mock.
  • F, G
    The ubiquitylation of PTEN was measured in (F) SGC7901 cells transfected with pcDNA‐CRNDE or Mock, and (G) 293T cells transfected with indicated vectors.

Source data are available online for this figure.