Journal: PLoS ONE

Article Title: Activation of p38/JNK Pathway Is Responsible for Embelin Induced Apoptosis in Lung Cancer Cells: Transitional Role of Reactive Oxygen Species

doi: 10.1371/journal.pone.0087050

Figure Lengend Snippet: ( A ) A549 cells were treated with 15 µM embelin for 4 and 8h. Total as well as phosphorylated ERK 1/2, p38, JNK 1/2 and tubulin (loading control) levels were measured by Western blot as described in the “Materials and Methods” section. ( B ) Normalized values of the band intensities obtained by densitometric analysis of the data from ( A ). ( C ) Caspase-3 activity in cells pre-treated with or without U0126 (5 µM), PD169316 (5 µM), SP600125 (5 µM) for 1h followed by embelin (15 µM) for 4h. Control values are normalized to 1 and the data indicated are the mean ± SD of three separate experiments. * indicates p<0.05 as compared with control and # indicates p<0.05 as compared with embelin treated cells.

Article Snippet: Blots were probed with monoclonal antibodies raised against total and phospho specific antibodies for ERK 1/2, p38, JNK (Cell Signaling Technology); and tubulin (Sigma-Aldrich).

Techniques: Western Blot, Activity Assay

Journal: PLoS ONE

Article Title: Activation of p38/JNK Pathway Is Responsible for Embelin Induced Apoptosis in Lung Cancer Cells: Transitional Role of Reactive Oxygen Species

doi: 10.1371/journal.pone.0087050

Figure Lengend Snippet: A549 cells were pre-treated with or without U0126 (5 µM), PD169316 (5 µM), SP600125 (5 µM) for 1h followed by embelin (15 µM) for 4h. Total and phosphorylated levels of ERK 1/2, p38, JNK 1/2 and tubulin (loading control) were detected by Western blotting as described in the “Materials and Methods” section.

Article Snippet: Blots were probed with monoclonal antibodies raised against total and phospho specific antibodies for ERK 1/2, p38, JNK (Cell Signaling Technology); and tubulin (Sigma-Aldrich).

Techniques: Western Blot

Journal: PLoS ONE

Article Title: Activation of p38/JNK Pathway Is Responsible for Embelin Induced Apoptosis in Lung Cancer Cells: Transitional Role of Reactive Oxygen Species

doi: 10.1371/journal.pone.0087050

Figure Lengend Snippet: A549 cells were pre-treated with or without the antioxidant FeTMPyP (10 µM) for 1h followed by embelin (15 µM) treatment for 4h. ( A ) Cellular levels of total and phosphorylated ERK 1/2, p38, JNK 1/2 and tubulin were detected by Western blot followed by chemiluminescence detection as described under “Materials and Methods” section. ( B ) Under similar experimental conditions as ( A ), cellular caspase-3 activity was measured as described under “Materials and Methods” section. Data presented are the mean ± SD of three separate experiments. * indicates p<0.01 as compared to control and # indicates p<0.05 as compared to embelin treated cells.

Article Snippet: Blots were probed with monoclonal antibodies raised against total and phospho specific antibodies for ERK 1/2, p38, JNK (Cell Signaling Technology); and tubulin (Sigma-Aldrich).

Techniques: Western Blot, Activity Assay

Journal: PLoS ONE

Article Title: Systemic Myostatin Inhibition via Liver-Targeted Gene Transfer in Normal and Dystrophic Mice

doi: 10.1371/journal.pone.0009176

Figure Lengend Snippet: C57 mice were injected intravenously with AAV 2/8 LSP.dnMstat and after one week the animals were sacrificed. Immunoblotting for total and phosphorylated Smad2/3, Akt and JNK was performed on protein homogenates of quadriceps muscle using beta-actin as a loading control. In dnMstat treated C57 quadriceps there was a (a) 41% decrease in phosphorylated Smad 2/3, (b) 59% increase in phosphorylated Akt and (c) 31% decrease in phosphorylated JNK. Data represent mean±SD. * Statistically significant compared to control, p<0.05.

Article Snippet: Se Jin Lee), phospho-Smad 2/3 (1∶500, Millipore, Temecula, CA), Smad 2/3 (1∶1000, Cell Signaling Technology, Danvers, MA), activin IIB receptor (1∶500, Sigma, St. Louis, MO), phospho-Akt and total Akt (1∶1000, Cell Signaling Technology, Danvers, MA), phospho-JNK and total JNK (1∶1000, Cell Signaling Technology, Danvers, MA), and actin (1∶2000, Sigma, St. Louis, MO).

Techniques: Injection, 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

Journal: Mediators of Inflammation

Article Title: Methimazole Inhibits the Expression of GFAP and the Migration of Astrocyte in Scratched Wound Model In Vitro

doi: 10.1155/2020/4027470

Figure Lengend Snippet: Methimazole attenuated the increased expression of phosphorylated JNK and ERK proteins (30 minutes). (a) Methimazole prevented the increased expression of p-ERK in scratched astrocytes. (b) Methimazole prevented the increased expression of p-JNK in scratched astrocytes. All values were expressed as mean ± SEM, n = 5. ∗ p < 0.05 vs. control; # p < 0.05 vs. scratch.

Article Snippet: Primary antibodies of GFAP (1 : 1000, Santa Cruz), p-ERK (1 : 1000, Cell Signaling), ERK (1 : 1000, Cell Signaling), p-JNK (1 : 1000, Cell Signaling), and JNK (1 : 1000, Cell Signaling) and the corresponding secondary antibodies were used here. β -Actin (Santa Cruz) was used as an internal control.

Techniques: Expressing