Fig. 4

SNHG1 promoted PTEN protein degradation. A and B Total RNA was extracted from the indicated UROtsa or U5637 transfectants using Trizol reagent. Real-Time PCR assays were then conducted to evaluate PTEN mRNA expression levels, utilizing GAPDH as an internal control. C and D The pMIR-PTEN 3’-UTR mRNA reporter was transiently transfected into the specified cells, followed by an assessment of the luciferase activity for each transfectant. The activity is depicted relative to scramble control transfectants and further normalized using an internal control, pRL-TK. Bars represent the mean ± SD from three separate experiments. E and F The designated cells were pretreated with the proteasome inhibitor MG132 (5 μM) for 10 h. Following this, the cells were exposed to CHX for the stipulated durations, and the cell extracts were subsequently analyzed through Western blotting. GAPDH served as a protein loading control