PTEN regulates pri-miR-21 processing.
(A) In vitro pri-miR-21 processing with Drosha-expressing cell lysates, Drosha immunoprecipitates, or glioblastoma cell lysates. Lane 1: probe only; lane 2: Drosha-WT overexpressed cell lysates; lane 3: Drosha-TN overexpressed cell lysates; lane 4: Drosha-WT immunoprecipitate; lane 5: Drosha-TN immunoprecipitate; lane 6: Drosha-WT immunoprecipitate; lane 7: LN428 cell lysates; lane 8: U87MG cell lysates. PTEN expression of LN428 (lane 9) and U87MG (lane 10) was confirmed by immunoblotting. (B) In vitro pri-miR-21 processing with parental 293T and PTEN expressing cell lysates. Lane 1: probe only; lane 2: parental 293T cell lysates; lane 3: PTEN expressing cell (clone #25) lysates; lane 4: PTEN expressing cell (clone #33) lysates. PTEN expression of parental 293T (lane 5) and PTEN expressing cells (lane 6: clone #25; lane 7: clone #33) was confirmed by immunoblotting. (C) In vitro pri-let-7a-1 processing with parental 293T and WT-PTEN expressing cell lysates. Lane 1: probe only; lane 2: parental 293T cell lysates; lane 3: PTEN expressing cell (clone #25) lysates; lane 4: PTEN expressing cell (clone #33) lysates. IB; immunoblot.