Journal: Experimental and Therapeutic Medicine

Article Title: Crosstalk between apoptosis and autophagy in prostate epithelial cells under androgen deprivation

doi: 10.3892/etm.2018.5726

Figure Lengend Snippet: Beclin-1 is cleaved during apoptosis. (A) Whole-cell lysates were prepared from androgen-deprived BPH-1 cells for 0, 12, 16, 20, 24, 28 and 32 h and analyzed by western blot analysis with anti-Beclin-1, anti-Caspase-3 and anti-GAPDH antibodies. (B) Relative expression of Caspase-3 proteins in BPH-1 cells under AD at 0 and 20 h was investigated using western blot analysis. Data are representative of the relative expression of proteins that were normalized by GAPDH. Data are presented as the mean ± standard deviation (n=3). **P<0.01. Beclin-1, a protein that in humans is encoded by the BECN1 gene; Caspase-3, cysteine-aspartic acid protease-3; AD, androgen deprivation.

Article Snippet: The primary antibodies used were as follows: PARP-1 polyclonal rabbit primary antibody (1:1,000); Caspase-3 primary antibody (1:1,000); LC-3 polyclonal rabbit primary antibody (1:1,000; all Cell Signaling Technology); Beclin-1 polyclonal rat primary antibody (1:2,000; BD Biosciences); and GAPDH polyclonal rabbit primary antibody (1:2,000; both BD Biosciences) as a reference protein.

Techniques: Western Blot, Expressing, Standard Deviation

Journal: Frontiers in Cellular Neuroscience

Article Title: Administration of Ketamine Causes Autophagy and Apoptosis in the Rat Fetal Hippocampus and in PC12 Cells

doi: 10.3389/fncel.2018.00021

Figure Lengend Snippet: Effects of ketamine on autophagy and apoptosis of the hippocampus in fetal rats. Increase in protein levels of C-Caspase-3 (16.4%) (A) , Beclin-1 (18.9%) (B) , Bax (16.1%) (C) , Atg5 (53.4%) (G) , and LC3-II (34.7%) (I) . Decrease in protein levels of Bcl-2 (23.2%) (D) , Atg4 (20.1%) (F) , p62 (36.9%) (H) , and LC3-I (40.3%) (I) The Bax/Bcl-2 ratio increased (E) , whereas the LC3-I/LC3-II ratio decreased (J) . * p < 0.05.

Article Snippet: Primary antibodies directed against LC3 (1.5:1,000; EnoGene, Nanjing, China), Cleaved-caspase 3 (1.5:1,000; EnoGene, Nanjing, China), Beclin-1 (1.5:1,000; EnoGene, Nanjing, China), Atg5 (1.5:1,000; EnoGene, Nanjing, China), Bax (1.5:1,000; EnoGene, Nanjing, China), Bcl-2 (1.5:1,000; EnoGene, Nanjing, China), Atg4 (1:1,000, Abcam, Cambridge, UK), or p62 (1:1,000; CST, Shanghai, China) were used in this study.

Techniques:

Journal: Frontiers in Cellular Neuroscience

Article Title: Administration of Ketamine Causes Autophagy and Apoptosis in the Rat Fetal Hippocampus and in PC12 Cells

doi: 10.3389/fncel.2018.00021

Figure Lengend Snippet: Effects of ketamine on PC12 cells. The protein levels of C-Caspase-3 (A) , Beclin-1 (B) , Bax (C) , Atg5 (G) , and LC3-II (I) increased after ketamine treatment, whereas Bcl-2 (D) , Atg4 (F) , p62 (H) , and LC3-I (I) decreased. The Bax/Bcl-2 ratio increased (E) and the LC3-I/LC3-II ratio decreased (J) after ketamine treatment. * p < 0.05.

Article Snippet: Primary antibodies directed against LC3 (1.5:1,000; EnoGene, Nanjing, China), Cleaved-caspase 3 (1.5:1,000; EnoGene, Nanjing, China), Beclin-1 (1.5:1,000; EnoGene, Nanjing, China), Atg5 (1.5:1,000; EnoGene, Nanjing, China), Bax (1.5:1,000; EnoGene, Nanjing, China), Bcl-2 (1.5:1,000; EnoGene, Nanjing, China), Atg4 (1:1,000, Abcam, Cambridge, UK), or p62 (1:1,000; CST, Shanghai, China) were used in this study.

Techniques:

Journal: Frontiers in Cellular Neuroscience

Article Title: Administration of Ketamine Causes Autophagy and Apoptosis in the Rat Fetal Hippocampus and in PC12 Cells

doi: 10.3389/fncel.2018.00021

Figure Lengend Snippet: Effects of ketamine on autophagy and apoptosis in PC12 cells. The protein levels of C-Caspase-3 (A) , Beclin-1 (B) , Bax (C) , Atg5 (G) , and LC3-II (I) increased after Rapa and ketamine treatment, whereas Bcl-2 (D) , Atg4 (F) , p62 (H) , and LC3-I (I) decreased. 3-MA resulted in accumulation of p62 (H) . In Rapa and K group, the values of log 2 Bax/Bcl-2 were significantly higher than those in C group (E) , while the values of Log 2 LC3-I/LC3-II were significantly lower than those in C group (J) . * p < 0.05.

Article Snippet: Primary antibodies directed against LC3 (1.5:1,000; EnoGene, Nanjing, China), Cleaved-caspase 3 (1.5:1,000; EnoGene, Nanjing, China), Beclin-1 (1.5:1,000; EnoGene, Nanjing, China), Atg5 (1.5:1,000; EnoGene, Nanjing, China), Bax (1.5:1,000; EnoGene, Nanjing, China), Bcl-2 (1.5:1,000; EnoGene, Nanjing, China), Atg4 (1:1,000, Abcam, Cambridge, UK), or p62 (1:1,000; CST, Shanghai, China) were used in this study.

Techniques:

Journal: Frontiers in Cellular Neuroscience

Article Title: Administration of Ketamine Causes Autophagy and Apoptosis in the Rat Fetal Hippocampus and in PC12 Cells

doi: 10.3389/fncel.2018.00021

Figure Lengend Snippet: Ketamine induces autophagy in PC12 cells by oxidative damage. Ketamine and H 2 O 2 induced changes in autophagy-related protein. The protein levels of C-Caspase-3 (A) , Beclin-1 (B) , Bax (C) , Atg5 (G) , and LC3-II (I) increased after H 2 O 2 and ketamine treatment, whereas Bcl-2 (D) , Atg4 (F) , p62 (H) , and LC3-I (I) decreased. In H 2 O 2 and K group, the values of log 2 Bax/Bcl-2 were significantly higher than those in C group (E) , while the values of Log 2 LC3-I/LC3-II were significantly lower than those in C group (J) . * p < 0.05.

Article Snippet: Primary antibodies directed against LC3 (1.5:1,000; EnoGene, Nanjing, China), Cleaved-caspase 3 (1.5:1,000; EnoGene, Nanjing, China), Beclin-1 (1.5:1,000; EnoGene, Nanjing, China), Atg5 (1.5:1,000; EnoGene, Nanjing, China), Bax (1.5:1,000; EnoGene, Nanjing, China), Bcl-2 (1.5:1,000; EnoGene, Nanjing, China), Atg4 (1:1,000, Abcam, Cambridge, UK), or p62 (1:1,000; CST, Shanghai, China) were used in this study.

Techniques:

Journal: Marine Drugs

Article Title: Biological Activities of Cyclic and Acyclic B-Type Laxaphycins in SH-SY5Y Human Neuroblastoma Cells

doi: 10.3390/md18070364

Figure Lengend Snippet: Analysis of p70 S6 kinase and Beclin 1 expression in neuroblastoma cells after treatment with B-type laxaphycins. The protein expression of ( a ) p70 S6 and ( b ) beclin 1 was determined by Western blot. Left panels show the results of compounds and right panels present the results of the positive controls bafilomycin A1 (Baf A1), rapamycin (Rap) and compound C (Comp C). The activation of the kinase was analyzed as the ratio between phosphorylated/total protein levels. Protein expression levels are normalized by β-actin. Values are mean ± SEM of three replicates performed in duplicate, expressed as percentage of control cells. * p < 0.05, ** p < 0.01

Article Snippet: LC3 II/I was detected with the primary antibody anti-LC3 II/I (1:1000, Abcam), the primary antibody anti-pmTOR (Ser2448) (1:1000, Merck Millipore) was used to recognize phospho-mTOR and the total levels of the kinase were determined with anti-mTOR antibody (1:10000, Abcam), beclin-1 was detected with anti-beclin-1 primary antibody (1:1000, Merck Millipore), p62 was quantified with anti-p62 antibody (1:1000, Merck Millipore), anti-phospho-p70 S6 kinase (Thr389) (1:1000, Merck Millipore) was used to detect phospho- p70 and the total levels were quantified using anti-p70 S6 kinase antibody (1:1000, Merck Millipore), anti-phospho-AMPK (Thr172) (1:500, Merck Millipore) was used to recognize the phosphorylated state of the kinase, and the total expression of the enzyme was detected with anti-AMPK antibody (1:500, Merck Millipore).

Techniques: Expressing, Western Blot, Activation Assay