Journal: Pharmaceutics

Article Title: Molecular Mechanisms Underlying the Anti-Inflammatory Properties of (R)-(-)-Carvone: Potential Roles of JNK1, Nrf2 and NF-κB

doi: 10.3390/pharmaceutics15010249

Figure Lengend Snippet: List of primary antibodies used in Western blot.

Article Snippet: SAPK/JNK , rabbit , polyclonal , 1:1000 , Cell Signaling Technology, Inc. , 9252/17.

Techniques: Western Blot

Journal: PLoS ONE

Article Title: Suppression of FOXM1 Sensitizes Human Cancer Cells to Cell Death Induced by DNA-Damage

doi: 10.1371/journal.pone.0031761

Figure Lengend Snippet: ( A ) MIA PaCa-2 control and FOXM1 knockdown pancreatic cancer cells were γ-irradiated with the indicated doses. Seventy-two hours following irradiation cells were harvested and immunoblotting was carried out with antibodies specific for Phospho/Total-SAPK/JNK and cleaved caspase-3. β-actin was used as the loading control. ( B ) MIA PaCa-2 FOXM1 knockdown pancreatic cancer cells were preincubated for 1 hour with 10 µM of JNK inhibitor, SP600125 and then were γ-irradiated as indicated. Forty-eight hours following irradiation cells were harvested and immunoblotting was performed for cleaved caspase-3 and β-actin as the loading control. ( C ) The graphs show mean values ± SEM of four independent experiments. ( D ) MDA-MB-231 FOXM1 knockdown breast cancer cells were preincubated for 1 hour with 10 µM of JNK inhibitor, SP600125 and then were subjected to γ-irradiation with the indicated doses. Seventy-two hours after irradiation cells were harvested and immunoblotting was performed with cleaved caspase-3 and β-actin antibodies. ( E ) The graphs show mean values ± SEM of two independent experiments. ( F ) Pancreatic MIA PaCa-2 control and FOXM1 knockdown cancer cells were subjected to ionizing radiation with the indicated doses. Thirty-six hours following irradiation cells were harvested and immunoblotting was carried out with antibodies specific for Bcl-2 and Bcl-xL. β-actin was used as the loading control. ( G ) Breast MDA-MB-231 control and FOXM1 knockdown cancer cells were γ-irradiated as indicated. Seventy-two hours following irradiation cells were harvested and immunoblotting was performed for Bcl-2. β-actin was used as the loading control. ( H ) MIA PaCa-2 control and FOXM1 knockdown pancreatic cancer cells were harvested for RNA extraction. Quantitative RT-PCR was carried out with Bcl-2 and cyclophilin primers. The graph demonstrates mean values ±SEM of three independent experiments. ( I ) To extract RNA MDA-MB-231 control and FOXM1 knockdown breast cancer cells were harvested. Using Bcl-2 and cyclophlin primers qRT-PCR was performed. The graph shows mean values ±SEM of three independent experiments.

Article Snippet: Treated cells were harvested and processed for immunoblotting as described in ref. with antibodies specific for FOXM1 (the rabbit polyclonal antibody against FOXM1 was described previously ), p53 (Santa Cruz), cleaved caspase-3 (Cell signaling), PARP-1/2 (Santa Cruz), Total and Phospho-SAPK/JNK (Cell signaling), Bcl-2 (Santa Cruz), Bcl-xL (Cell signaling) and β-actin (Sigma).

Techniques: Irradiation, Western Blot, RNA Extraction, Quantitative RT-PCR