Journal: Cell Stress & Chaperones

Article Title: Low-pH-induced apoptosis: role of endoplasmic reticulum stress-induced calcium permeability and mitochondria-dependent signaling

doi: 10.1007/s12192-014-0568-6

Figure Lengend Snippet: Acidic stress-induced apoptosis in Raji cells. a Percentage viability of Raji cells grown in media of pH 7.3, 6.8, 6.3, and 5.8 after 12, 24, and 48 h. b Detection of apoptosis by DNA laddering assay. Raji cells were grown in acidic environment (pH 6.8, pH 6.3, and pH 5.8) for 48 h, and the cells grown at pH 7.3 were taken as control

Article Snippet: Exponentially growing Raji cells (human acute lymphoid leukemia cells) were procured from the National Centre for Cell Science (NCCS), Pune.

Techniques: DNA Laddering, Control

Journal: Cell Stress & Chaperones

Article Title: Low-pH-induced apoptosis: role of endoplasmic reticulum stress-induced calcium permeability and mitochondria-dependent signaling

doi: 10.1007/s12192-014-0568-6

Figure Lengend Snippet: Increase in intracellular calcium at acidic pH. Raji cells were cultured in normal (pH 7.3) and acidic media pH 6.8 and pH 5.8 for 48 h followed by incubation with the fluorescent dye Fluo 3-AM. Representative FACS analysis histograms of Fluo 3-AM stained Raji cells in control and after acidic stress treatment. Five thousand events were counted. Increase in calcium at pH 5.8 was found statically significant (p value <0.005) with respect to control. Cells at physiological pH (pH 7.3) were taken as control

Article Snippet: Exponentially growing Raji cells (human acute lymphoid leukemia cells) were procured from the National Centre for Cell Science (NCCS), Pune.

Techniques: Cell Culture, Incubation, Staining, Control

Journal: Cell Stress & Chaperones

Article Title: Low-pH-induced apoptosis: role of endoplasmic reticulum stress-induced calcium permeability and mitochondria-dependent signaling

doi: 10.1007/s12192-014-0568-6

Figure Lengend Snippet: Effect of acidic stress on XBP1 splicing, CHOP, and GRP78 in Raji cells. Raji cells were grown at pH 7.3, pH 6.8, and pH 5.8 (acidic stress) for 48 h. Total RNA was extracted from the cells, and the mRNA of XBP1 was detected and analyzed by RT-PCR. The RT-PCR products of XBP1 were analyzed on 12.5 % PAGE, and GRP78 and CHOP were analyzed on 2 % agarose gel. β-Actin was used as internal control. Cells grown at physiological pH 7.3 were taken as control

Article Snippet: Exponentially growing Raji cells (human acute lymphoid leukemia cells) were procured from the National Centre for Cell Science (NCCS), Pune.

Techniques: Reverse Transcription Polymerase Chain Reaction, Agarose Gel Electrophoresis, Control

Journal: Cell Stress & Chaperones

Article Title: Low-pH-induced apoptosis: role of endoplasmic reticulum stress-induced calcium permeability and mitochondria-dependent signaling

doi: 10.1007/s12192-014-0568-6

Figure Lengend Snippet: Bax induction and translocation to mitochondrion by low pH. a Raji cells were incubated in acidic environment pH 6.8 and pH 5.8 for 48 h. Expression analysis was carried out by quantitative real-time PCR. Real-time PCR was carried out using gene-specific primers, and specificity of product was confirmed by melting curve analysis. The data was normalized to β-actin. b Subcellular fractionation was performed to obtain the fractions for cytosol, mitochondria, and ER/microsome followed by western blot analysis using anti Bax-specific monoclonal antibody. Under acidic environment, a significant increase in Bax expression at pH 5.8 was observed in mitochondrion fraction while in contrast to it, Bax expression decreased in endoplasmic fractions. Cells grown at physiological pH (pH 7.3) were taken as control

Article Snippet: Exponentially growing Raji cells (human acute lymphoid leukemia cells) were procured from the National Centre for Cell Science (NCCS), Pune.

Techniques: Translocation Assay, Incubation, Expressing, Real-time Polymerase Chain Reaction, Fractionation, Western Blot, Control

Journal: Cell Stress & Chaperones

Article Title: Low-pH-induced apoptosis: role of endoplasmic reticulum stress-induced calcium permeability and mitochondria-dependent signaling

doi: 10.1007/s12192-014-0568-6

Figure Lengend Snippet: Acidic stress treatment leads to enhanced cytochrome c release and caspase-3 activation. a Representative FACS analysis histograms of anti-cytochrome c stained Raji cells after acidic stress. Bar graph showing cytochrome c expression after acidic stress treatment expressed as mean florescence intensity. b Representative FACS analysis histograms of anti-active caspase-3 stained Raji cells after acidic stress. Bar graph showing caspase-3 expressions after acidic stress treatment expressed as mean florescence intensity. A significant (P value <0.005) upregulation in active caspase-3 was observed with respect to control. Cells grown at physiological pH (pH 7.3) were taken as control. Five thousand events were counted per tube. c Illustrated PARP cleavage leads to generation of 85-kDa fragments as compare to control

Article Snippet: Exponentially growing Raji cells (human acute lymphoid leukemia cells) were procured from the National Centre for Cell Science (NCCS), Pune.

Techniques: Activation Assay, Staining, Expressing, Control

Journal: Cell Stress & Chaperones

Article Title: Low-pH-induced apoptosis: role of endoplasmic reticulum stress-induced calcium permeability and mitochondria-dependent signaling

doi: 10.1007/s12192-014-0568-6

Figure Lengend Snippet: Low pH caused ROS independent cell death. Raji cells exposed to acidic stress were incubated with DCFH-DA after and 48 h followed by flow cytometry analysis. Cells grown at physiological pH (pH 7.3) were taken as control. Acidic stress-mediated induction of apoptosis was found to be independent of ROS

Article Snippet: Exponentially growing Raji cells (human acute lymphoid leukemia cells) were procured from the National Centre for Cell Science (NCCS), Pune.

Techniques: Incubation, Flow Cytometry, Control

Journal: Cell Stress & Chaperones

Article Title: Low-pH-induced apoptosis: role of endoplasmic reticulum stress-induced calcium permeability and mitochondria-dependent signaling

doi: 10.1007/s12192-014-0568-6

Figure Lengend Snippet: Acidic environment upregulated mRNA level of TP53 and p21 and elevated the expression of NF-κB, p53, and p21 in Raji cells. a Quantitative real-time PCR-based analysis of TP53 and p21 expression. RNA was isolated from control and acidic stress (pH 5.8 and pH 6.8) exposed Raji cells. Real-time PCR was carried out using gene-specific primers, and specificity of product was confirmed by melting curve analysis. A significant (P value <0.005) increase in p21 mRNA was found at pH 6.8 and pH 5.8 with respect to control. b Representative FACS analysis histograms of anti-p21 stained Raji cells after acidic stress. Bar graph showing p21 expression after acidic stress treatment expressed as mean florescence intensity. c Induction of p53 and nuclear localization of NF-κB by acidic environment in Raji cells. The data was normalized to β-actin for real-time analysis and GAPDH for Western blot analysis. Cells at pH (pH 7.3) were taken as control

Article Snippet: Exponentially growing Raji cells (human acute lymphoid leukemia cells) were procured from the National Centre for Cell Science (NCCS), Pune.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Isolation, Control, Staining, Western Blot

Journal: PLoS ONE

Article Title: A Novel BAT3 Sequence Generated by Alternative RNA Splicing of Exon 11B Displays Cell Type-Specific Expression and Impacts on Subcellular Localization

doi: 10.1371/journal.pone.0035972

Figure Lengend Snippet: HeLa cells (Adenocarcinoma), the human melanoma cell line MelJuSo, the B lymphoma Raji and monocyte-derived dendritic cells (moDCs) were plated on coverslips and stained for BAT3 using a polyclonal serum against a C-terminal peptide (middle lane). Cell nuclei (left lane) were visualized with DAPI (A) or 7AAD (B). Merged images are shown in the right lane. A. Immunofluorescence staining was evaluated with a standard fluorescence microscope and B. by confocal microscopy. Scale bars = 10 µm. C. Nuclear and cytosolic staining of endogenous BAT3 in Raji cells was evaluated in 10 single cells using ImageJ. MFI, mean fluorescence intensity per region of interest D. Western blot analysis of subcellular fractions from Raji and HeLa cells. Nuclei (N) and cytoplasm (C) were separated by SDS-PAGE and immunoblotted for BAT3, GADPH (cytosolic marker) and histone H3 (nuclear marker).

Article Snippet: The B lymphoma cell line Raji was cultured in RPMI and dendritic cells were grown in VLE-RPMI supplemented with antibiotics and glutamine (Biochrom AG, Berlin, Germany).

Techniques: Derivative Assay, Staining, Immunofluorescence, Fluorescence, Microscopy, Confocal Microscopy, Western Blot, SDS Page, Marker

Journal: PLoS ONE

Article Title: A Novel BAT3 Sequence Generated by Alternative RNA Splicing of Exon 11B Displays Cell Type-Specific Expression and Impacts on Subcellular Localization

doi: 10.1371/journal.pone.0035972

Figure Lengend Snippet: A. HeLa cells transfected with BAT3 Δ11B,24 were cultured on coverslips in the presence (lower panel) or absence (upper panel) of leptomycin B (LMB) for 2 h. Cells were subsequently stained with V5 mAb and inspected with standard immunofluorescence microscopy (second panel). Left panel shows DAPI stained nuclei, third panel merging of images and right panel displays phase contrast images. Scale bars = 10 µm. B. Raji cells were cultured for 2 h in the presence (lower panel) or absence (upper panel) of LMB and then plated on coverslips. Cells were subsequently stained with the polyclonal anti-BAT3 serum and with ISCR3 mAb (HLA-DR) for evaluation by immunofluorescence microscopy. Left panel shows DAPI staining, second panel staining for BAT3, third panel staining for HLA-DR and images were merged in the right panel. Scale bars = 5 µm.

Article Snippet: The B lymphoma cell line Raji was cultured in RPMI and dendritic cells were grown in VLE-RPMI supplemented with antibiotics and glutamine (Biochrom AG, Berlin, Germany).

Techniques: Transfection, Cell Culture, Staining, Immunofluorescence, Microscopy

Journal: Frontiers in Medicine

Article Title: Development of combinatorial antibody therapies for diffuse large B cell lymphoma

doi: 10.3389/fmed.2022.1034594

Figure Lengend Snippet: Antibodies induce ADCC and ADCP responses within lymphoma cell lines. (A) Schematic representation of the assays used to measure ADCC and ADCP response within modified Jurkat effector cells. Created with BioRender.com . (B) Line graphs of the relative fluorescent units (RFU) of luciferase produced by ADCC activation of FcγRIIIa modified Jurkat cells when incubated with Raji, SU-DHL-4, NU-DUL-1, or SU-DHL-8 DLBCL cell lines and serial diluted anti-CD20 (Blue), bispecific anti-CD20/anti-CD74 (Purple), or bispecific anti-CD20/anti-IL4R (Orange) antibodies. (C) Line graph of the relative fluorescent units (RFU) of luciferase produced by ADCC activation of FcγRIIa-H modified Jurkat cells when incubated with Burkitt’s lymphoma Raji cells and serial diluted anti-CD20 (Blue), bispecific anti-CD20/anti-CD74 (Purple), or bispecific anti-CD20/anti-IL4R (Orange) antibodies.

Article Snippet: ADCP Bioassay Complete Kit (Promega) was performed using the manufacturer protocol with the provided Raji target cells and modified Jurkat NFAT-luc FcγRIIa-H effector cells.

Techniques: Modification, Luciferase, Produced, Activation Assay, Incubation

Journal: Frontiers in Oncology

Article Title: Tissue Factor-Targeted “O 2 -Evolving” Nanoparticles for Photodynamic Therapy in Malignant Lymphoma

doi: 10.3389/fonc.2020.524712

Figure Lengend Snippet: The antitumor effect and safety evaluation in vivo . Sections (A–E) correspond to the Raji mouse model. (A) PET imaging performed 24 h before and 24 h after PDT treatment. (B) Quantitative analysis of tumor growth inhibition rate by mean radioactivity of 18F-FDG. *p < 0.05, ***p < 0.001. (C) Relative tumor volume variation of mice treated under different conditions. (D) H&E staining and TUNEL staining of tumors at 24 h after PDT treatment. Apoptotic cells were identified by TUNEL assay (brown) (scale bar 100 μm). (E) H&E stained images of major organ slices (scale bar 200 μm). (F) Tumors at 24 h post-treatment and corresponding TUNEL staining of tumor slides in the Jeko-1 mouse model. (scale bar 50 μm).

Article Snippet: Cells were maintained in RPMI-1640 supplemented with 10% FBS; the Jeko-1 and Raji cell lines were purchased from the China Center for Type Culture Collection (Wuhan University, China) and cultured in an RPMI-1640 culture medium containing 10% FBS at 37°C in a humidified atmosphere enclosing 5% CO 2 .

Techniques: In Vivo, Imaging, Inhibition, Radioactivity, Staining, TUNEL Assay