Journal: PLoS ONE

Article Title: JNK and Macroautophagy Activation by Bortezomib Has a Pro-Survival Effect in Primary Effusion Lymphoma Cells

doi: 10.1371/journal.pone.0075965

Figure Lengend Snippet: (A) Time course assay was performed on BC3 and BCBL1 cells treated with 20 nM of bortezomib (BZ) for 0, 4, 8, 16 hrs. Total cell lysates were prepared and immunoblotted with anti-BiP, anti-CHOP, anti IRE1α, anti-phospho-JNK (p-JNK 1/2) and t-JNK antibodies. Anti-tubulin was assayed as protein loading control. (B) Electron microscopy (EM) analysis was performed on BC3 and BCBL1 cells treated with Bortezomib (20 nM) for 16 hrs (BZ). Cells showed several signs of ER stress compared to untreated control (CT). More apoptotic features such as nuclear (N) condensation were observed in BCBL1. Er (Endoplasmic reticulum), G (Golgi Apparatus), N (Nucleus), Au (Autophagosomes), Ne (Nuclear Envelope), M (Mitochondria). Bars: 1 µm. Results are representative of three independent experiments.

Article Snippet: The BC3, BCBL1, JSC1 and BC1 PEL cell lines (ATCC) were cultured in RPMI 1640 10% Fetal Calf Serum (FBS) (Euroclone), glutamine and streptomycin (100 μg/ml) and penicillin (100U/ml) in 5% CO 2 at 37°C.

Techniques: Control, Electron Microscopy

Journal: PLoS ONE

Article Title: JNK and Macroautophagy Activation by Bortezomib Has a Pro-Survival Effect in Primary Effusion Lymphoma Cells

doi: 10.1371/journal.pone.0075965

Figure Lengend Snippet: (A) Electron microscopy (EM) on BC3 and BCBL1 cells treated with 20 nM of bortezomib after 16 hrs shows pronounced cell vacuolization (A a, a 4 ) with autophagosome membrane-bound compartment in different stage of maturation (A a 1, a 2 , a 4, a 5 ). Small intracellular structures consistent with aggresomes were evident in BCLB1 cells (A a 6 ). N (Nucleus), Au (Autophagosomes), M (Mitochondria), ag (aggresomes). Bars 1 µm. Results are representative of three independent experiments. (B) PEL cells were treated with bortezomib (20 nM), bafilomycin A (Baf) (10 nM) or combination of both. The western blot analysis was performed to detect the two isoforms of LC3 protein. The histograms indicate LC3-II/I and LC3-II/Actin ratio based on densitometric analysis (mean ± the standard deviation, n = 3 experiments). An antibody against β-actin was used as loading control.

Article Snippet: The BC3, BCBL1, JSC1 and BC1 PEL cell lines (ATCC) were cultured in RPMI 1640 10% Fetal Calf Serum (FBS) (Euroclone), glutamine and streptomycin (100 μg/ml) and penicillin (100U/ml) in 5% CO 2 at 37°C.

Techniques: Electron Microscopy, Membrane, Western Blot, Standard Deviation, Control

Journal: PLoS ONE

Article Title: JNK and Macroautophagy Activation by Bortezomib Has a Pro-Survival Effect in Primary Effusion Lymphoma Cells

doi: 10.1371/journal.pone.0075965

Figure Lengend Snippet: (A) BC3 and BCBL1 cells were treated with bortezomib (20 nM for 16 hrs) with or without pre-treatment with JNK inhibitor (SP600125) (20 µM). Total cell lysates were prepared and immunoblotted with the following antibodies: anti-LC3, anti-pJNK, anti-T-JNK and anti-p62. Anti-β-actin was used as loading control. The histograms indicate LC3-II/I and LC3-II/Actin ratio based on densitometric analysis (mean ± the standard deviation, n = 3 experiments). (B) BC3 cells were transfected with DN-JNK expression vector or with an control empty vector (CV) and after 16 hrs treated with bortezomib (20 nM) for an additional 16 hrs. Total cell lysates were prepared and immunoblotted with anti-LC3 and p62 antibodies. β-actin was used as internal control. The histograms indicate LC3-II/I and LC3-II/Actin ratio based on densitometric analysis (mean ± the standard deviation, n = 3 experiments). (C) BC3 and BCBL1 cells were treated with Bortezomib (20 nM) alone or in combination with JNK inhibitor (SP600125) (20 µM) or ERK inhibitor (PD98059) (10 µM) for 16 hrs. A western blotting was performed using the following antibodies: anti-pBcl2(S70) and anti-total Bcl2. β-actin was used as loading control.

Article Snippet: The BC3, BCBL1, JSC1 and BC1 PEL cell lines (ATCC) were cultured in RPMI 1640 10% Fetal Calf Serum (FBS) (Euroclone), glutamine and streptomycin (100 μg/ml) and penicillin (100U/ml) in 5% CO 2 at 37°C.

Techniques: Control, Standard Deviation, Transfection, Expressing, Plasmid Preparation, Western Blot

Journal: PLoS ONE

Article Title: JNK and Macroautophagy Activation by Bortezomib Has a Pro-Survival Effect in Primary Effusion Lymphoma Cells

doi: 10.1371/journal.pone.0075965

Figure Lengend Snippet: (A) Viability assay evaluated by trypan blue exclusion was performed in BC3 (left panel) and BCBL1 (right panel) cells treated with bortezomib (20 nM) or 3-MA (5 mM) alone or in combination for 16 hrs. Mean ± the standard deviation was also indicated (n = 3 experiments). * p-value = 0.05, ** p-value = 0.02. Western blotting analysis was performed on BC3 cells to evaluate the expression of cleaved (cl) PARP p85 fragment (middle panel). β-actin was used as loading control. (B) BC3 and BCBL1 cells were transfected with ATG5 siRNA or scramble siRNA (siRNASc), and than a western blot was performed with the anti-ATG5 antibody. β-actin was used as loading control. Viability assay evaluated by trypan blue exclusion was performed in BC3 and BCBL1 cells ATG5 or scramble-knocked down upon bortezomib treatment (20 nM) for 16 hrs. Mean ± the standard deviation was indicated (n = 3 experiments). ♦ p-value = 0.02, ♦♦ p-value = 0.03. Western blotting analysis was also performed on BC3 cells to evaluate the expression of PARP p85 fragment (cl PARP) and β-actin was used as internal control. (C) Cells viability assay on BC3 and BCBL1 cells treated with bortezomib (20 nM) and SP600125 (20 µM) alone or in combination for 16 hrs. The percentage of live cells was evaluated by trypan blue exclusion assay. Mean ± the standard deviation was indicated (n = 3 experiments). p-value = 0.01, p-value = 0.01. Western blotting analysis was performed on BC3 cells to evaluate the expression of PARP p85 fragment (cl PARP) and β-actin was used as loading control.

Article Snippet: The BC3, BCBL1, JSC1 and BC1 PEL cell lines (ATCC) were cultured in RPMI 1640 10% Fetal Calf Serum (FBS) (Euroclone), glutamine and streptomycin (100 μg/ml) and penicillin (100U/ml) in 5% CO 2 at 37°C.

Techniques: Viability Assay, Standard Deviation, Western Blot, Expressing, Control, Transfection, Trypan Blue Exclusion Assay

Journal: Cell reports

Article Title: KSHV episome tethering sites on host chromosomes and regulation of latency-lytic switch by CHD4

doi: 10.1016/j.celrep.2022.110788

Figure Lengend Snippet:

Article Snippet: Human: BC3 , ATCC , CRL-2277.

Techniques: Produced, Virus, Recombinant, Protease Inhibitor, Magnetic Beads, Luciferase, Sequencing, In Vitro, Software

Journal: Oxidative medicine and cellular longevity

Article Title: Selenium Attenuates TBHP-Induced Apoptosis of Nucleus Pulposus Cells by Suppressing Mitochondrial Fission through Activating Nuclear Factor Erythroid 2-Related Factor 2.

doi: 10.1155/2022/7531788

Figure Lengend Snippet: Figure 2: Se ameliorated TBHP-induced apoptosis and ECM degeneration of NPCs. (a, b) Apoptosis of the NPCs detected by flow cytometry analysis. (c, d) Typical fluorescence photomicrograph and quantitation of TUNEL staining of NPCs (scale bar: 100 μm). (e, f) Representative western blot bands and quantitation of the expression of cleaved-caspase 9, cleaved-caspase 3, Bax, and Bcl-2 in the NPCs. (g, h) Representative western blot bands and quantitation of the expression of anabolic mediators (aggrecan and collagen II) and catabolic mediators (MMP3 and MMP9). The data are represented as the mean ± SD from at least 3 independent experiments. ∗P < 0:05 and ∗∗P < 0:01 vs. the TBHP group.

Article Snippet: After being blocked with 5% skimmed milk for 1 h at room temperature, the PVDF membranes were incubated overnight at 4°C with the primary antibodies against Nrf2 (1 : 1000, ProteinTech, Wuhan, China), HO-1 (1 : 1000, ProteinTech), SOD2 (1 : 2000, ProteinTech), dynamin-related protein 1 (DRP1) (1 : 1000, Affinity Biosciences, OH, USA), mitochondrial fission factor (MFF) (1 : 1000, ProteinTech), mitochondrial fission 1 (Fis1) (1 : 1000, ProteinTech), optic atrophy 1 (OPA1) (1 : 1000, ProteinTech), mitofusin 1 (Mfn1) (1 : 1000, ProteinTech), mitofusin 2 (Mfn2) (1 : 1000, ProteinTech), Aggrecan (1 : 500, NOVUS, USA), collagen II (1 : 1000, Affinity Biosciences), matrix metalloproteinase 3 (MMP3) (1 : 1000, Affinity Biosciences), MMP9 (1 : 1000, Affinity Biosciences), cleaved-caspase 3 (1 : 1000, Affinity Biosciences), caspase 9 (1 : 1000, ProteinTech), Bcl-2 (1 : 1000, Abcam, Cambridge, MA, USA), Bax (1 : 1000, ProteinTech), and β-actin (1 : 2000, Cell Signaling Technology, Danvers, MA, USA).

Techniques: Cytometry, Quantitation Assay, TUNEL Assay, Staining, Western Blot, Expressing

Journal: Antibody Therapeutics

Article Title: Biology drives the discovery of bispecific antibodies as innovative therapeutics

doi: 10.1093/abt/tbaa003

Figure Lengend Snippet: Programs in clinical and preclinical stages to block the angiogenesis and/or tumorigenesis for cancer treatment

Article Snippet: TXB4-BC1 , Ossianix Inc , TfR × CD20 , Preclinical , Anti-tumorigenesis , Trojan horse , Fab + SDA with Fc, 2 + 2 , NA.

Techniques: Blocking Assay

Journal: Antibody Therapeutics

Article Title: Biology drives the discovery of bispecific antibodies as innovative therapeutics

doi: 10.1093/abt/tbaa003

Figure Lengend Snippet: Programs in clinical and preclinical stages to enhance tumor immunity for cancer treatment

Article Snippet: TXB4-BC1 , Ossianix Inc , TfR × CD20 , Preclinical , Anti-tumorigenesis , Trojan horse , Fab + SDA with Fc, 2 + 2 , NA.

Techniques: