Fig. 5: AMPK activation mediates ASP4132-induced anti-NSCLC cell activity.
From: AMPK activation by ASP4132 inhibits non-small cell lung cancer cell growth
Stable pNSCLC-1 cells, expressing the lentiviral AMPKα1 shRNA (“shAMPKα1”), the CRISPR/Cas9-AMPKα1-KO construct (“koAMPKα1”) (A–D), the dominate negative AMPKα1 (T172A, “dnAMPKα1”) (E–H), the constitutively active AMPKα1 (T172D, caAMPKα1) construct (I–L), or corresponding control shRNA or empty construct, were established; Cells were treated with ASP4132 (1 μM) or vehicle control and cultured for applied time periods, expression of listed proteins was tested by Western blotting assays (A, E, I). Cell viability, apoptosis and death were tested by CCK-8 (B, F, J), apoptotic nuclei staining (C, G, K) and Trypan blue staining (D, H, L) assays, respectively. Expression of listed proteins was quantified and normalized to the loading control (A, E, I). Red stars indicated expression of the mutant AMPKα1 (E, I). “Veh” stands for the vehicle control (Saline). Data were presented as mean ± standard deviation (SD, n = 5). “sh-C + Cas9-C” stands for control pNSCLC-1 cells with scramble control shRNA plus CRISPR/Cas9 empty vector (A–D). “Vec” stands for empty vector (E–L). *p < 0.05 vs. “Veh” cells. #p < 0.05 vs. ASP4132-treatment in “sh-C + Cas9-C” control cells or “Vec” control cells. “n.s.” stands for non-statistical difference. The experiments were repeated five times with similar results detected.
