Journal: Oncotarget

Article Title: Targeted activation of AMPK by GSK621 ameliorates H 2 O 2 -induced damages in osteoblasts

doi: 10.18632/oncotarget.14454

Figure Lengend Snippet: Neomycin-selected stable MC3T3-E1 cells expressing dominant negative AMPKα1 (T172A) construct (“dn-AMPKα1”, Flag-tagged) or the empty vector (pSuper-Flag, “Vec”) were treated with GSK621 (10 μM) for 2 hours, expression of listed proteins was tested A . Cells were also subjected to H 2 O 2 (250 μM) stimulation; Cell survival (CCK-8 assay, B . Cell death (LDH release assay, C . and apoptosis (Histone-DNA ELISA assay, D . were tested. Data are shown as mean (n=4) ± SD. p-AMPKα1 and p-ACC were quantified. Experiments in this figure were repeated for three times, and similar results were obtained. # p <0.05 vs. “Vec” group.

Article Snippet: The two non-overlapping murine AMPKα1 siRNAs (“Seq-1/Seq-2”) and the scramble control siRNA were synthesized by Genepharm.

Techniques: Expressing, Dominant Negative Mutation, Construct, Plasmid Preparation, CCK-8 Assay, Lactate Dehydrogenase Assay, Enzyme-linked Immunosorbent Assay

Journal: Oncotarget

Article Title: Targeted activation of AMPK by GSK621 ameliorates H 2 O 2 -induced damages in osteoblasts

doi: 10.18632/oncotarget.14454

Figure Lengend Snippet: MC3T3-E1 cells were treated with GSK621 at applied concentrations for 2 hours, expression of listed proteins was shown A . MC3T3-E1 cells, expressing listed lentiviral AMPKα1 shRNA (“Seq-1/-2/3”) or scramble control shRNA (“sc-sh”), were subjected to Western blotting assay of listed proteins B .; These cells were also treated with/out H 2 O 2 (250 μM) or plus GSK621 (10 μM) for 24 hours; Cell viability (CCK-8 assay, C . and D .) and cell death (LDH release assay, E . were tested. Primary murine osteoblasts, transfected with indicated AMPKα1 siRNA (“Seq-1/-2”) or scramble control siRNA (“sc-si”), were treated with GSK621 (10 μM) for 2 hours, expression of listed proteins was tested F . Cells were also stimulated with H 2 O 2 (250 μM) for 24 hours, and cell survival G . and cell death H . were tested. AMPKα1 (“p-” and total) or p-ACC were quantified. Data are shown as mean (n=6) ± SD. Experiments in this figure were repeated for three times, and similar results were obtained. # p <0.05 vs. “sc-sh”/“sc-si” group.

Article Snippet: The two non-overlapping murine AMPKα1 siRNAs (“Seq-1/Seq-2”) and the scramble control siRNA were synthesized by Genepharm.

Techniques: Expressing, shRNA, Control, Western Blot, CCK-8 Assay, Lactate Dehydrogenase Assay, Transfection

Journal: Oncotarget

Article Title: Targeted activation of AMPK by GSK621 ameliorates H 2 O 2 -induced damages in osteoblasts

doi: 10.18632/oncotarget.14454

Figure Lengend Snippet: Puromycin-selected stable MC3T3-E1 cells, expressing the constitutively-active AMPKα1 (T172D) construct (“ca-AMPKα1”, Flag-tagged) or the empty vector (pSuper-Flag, “Vector”), were subjected to Western blotting assay of listed proteins A . Above cells were also treated with H 2 O 2 (250 μM) or plus GSK621 (10 μM) for 24 hours; Cell survival (CCK-8 assay, B . and cell death (LDH release assay, C . were tested. Data are shown as mean (n=4) ± SD. p-AMPKα1 and p-ACC were quantified. Experiments in this figure were repeated for four times, and similar results were obtained. # p <0.05 vs. “Vector” group.

Article Snippet: The two non-overlapping murine AMPKα1 siRNAs (“Seq-1/Seq-2”) and the scramble control siRNA were synthesized by Genepharm.

Techniques: Expressing, Construct, Plasmid Preparation, Western Blot, CCK-8 Assay, Lactate Dehydrogenase Assay

Journal: Oncotarget

Article Title: Targeted activation of AMPK by GSK621 ameliorates H 2 O 2 -induced damages in osteoblasts

doi: 10.18632/oncotarget.14454

Figure Lengend Snippet: MC3T3-E1 cells, expressing scramble control shRNA (“sc-sh”), AMPKα1 shRNA (Seq-1, “sh AMPKα1”) or dominant negative AMPKα1 (T172A) (“dn-AMPKα1”), were treated with GSK621 (10 μM) or plus H 2 O 2 (250 μM) for applied time; Relative NAPDH level A . and ROS content B . were tested by the listed assays. The primary murine osteoblasts were treated with GSK621 (10 μM) or plus H 2 O 2 (250 μM) for applied time, relative NAPDH level C . and ROS content D . were tested. Data are shown as mean (n=3) ± SD. Experiments in this figure were repeated for four times, and similar results were obtained. * p <0.05 vs. “Ctrl” group. # p <0.05 vs. “H 2 O 2 ” only group.

Article Snippet: The two non-overlapping murine AMPKα1 siRNAs (“Seq-1/Seq-2”) and the scramble control siRNA were synthesized by Genepharm.

Techniques: Expressing, Control, shRNA, Dominant Negative Mutation