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