Molecules and Cells

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Fig. 4.

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Fig. 4. (A and B) C3H10T1/2 cells were transfected with pcDNA3.0-AMPK (2 μg) for 6 h and then incubated for a further 12 h. (A) RT-PCR analysis was performed using total RNA extracted from cells. (B) Western blot analysis was performed using the indicated antibodies. (C and D) C3H10T1/2 cells were treated with BMP2 (0.25 μg/ml) for 24 h. Com.C (+, 0.1 μM; ++, 1 μM) was added 3 h prior to harvest. (C) RT-PCR analysis was performed using total RNA extracted from cells. (D) Western blot analysis was performed using the indicated antibodies. (E) C3H10T1/2 cells were transiently transfected with pc-DNA3.0AMPK (0.4 μg/well) and/or treated with BMP2 (0.25 μg/ml) for 4 days, and then stained with BCIP/NBT liquid substrate. **P < 0.001 compared with untreated control cells. #P < 0.05 compared with CA-AMPK transfected cells. (F) C3H10T1/2 cells were treated with or without BMP2 (0.25 μg/ml) in the presence of Com.C (1 μM) for 4 days and then ALP staining was performed. *P < 0.05 compared with untreated control cells. #P < 0.05 compared with BMP2 treated cells. (G) C3H10T1/2 cells were transfected with FTO-Luc (2 μg), pc-DNA3.0-AMPK (2 μg) and b-galactosidase plasmids. After 24 h, cultures were treated with BMP2 (0.25 μg/ml) or Com.C (1 μM) for 48 h. Luciferase activity was measured and presented as the mean ± SEM of individual experiments. ***P < 0.005 compared with untreated control cells. &&&P < 0.005 compared with BMP2 treated cells. ###P < 0.005 compared with pc-DNA-3.0-AMPK-transfected cells. All experiments were independently repeated at least three times.
Mol. Cells 2020;43:58~65 https://doi.org/10.14348/molcells.2019.0136
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