Journal: iScience

Article Title: Identification of lysine-lactylated substrates in gastric cancer cells

doi: 10.1016/j.isci.2022.104630

Figure Lengend Snippet:

Article Snippet: MGC-803 cells , iCell Bioscience , iCell-h141.

Techniques: Control, Recombinant, Magnetic Beads, CCK-8 Assay, Mass Spectrometry, Software

Journal: Analytical Cellular Pathology (Amsterdam)

Article Title: TWIST1-EP300 Expedites Gastric Cancer Cell Resistance to Apatinib by Activating the Expression of COL1A2

doi: 10.1155/2022/5374262

Figure Lengend Snippet: The COL1A2 promoter is significantly regulated by an enhancer-like signature. (a) COL1A2 promoter modification level searched by UCSC browser. (b, c) ChIP-seq data of H3K27ac in GC and normal gastric tissues were downloaded from the GEO database and analyzed for enrichment in the COL1A2 promoter. (d) ChIP-qPCR analysis of COL1A2 promoter histone H3K27ac modification levels in AGS and MKN-45 cells. Data indicate means ± SD of three biological replicates. Two-way ANOVA and Tukey's multiple comparison test; ∗∗ p < 0.01 (vs. IgG).

Article Snippet: Human GC cell lines MKN-45 and AGS were purchased from China Center for Type Culture Collection (Wuhan, Hubei, China).

Techniques: Modification, ChIP-sequencing

Journal: Analytical Cellular Pathology (Amsterdam)

Article Title: TWIST1-EP300 Expedites Gastric Cancer Cell Resistance to Apatinib by Activating the Expression of COL1A2

doi: 10.1155/2022/5374262

Figure Lengend Snippet: EP300 promotes COL1A2 expression through an H3K27ac-dependent manner. GC cells were treated with HAT inhibitor or DMSO. (a) ChIP-qPCR analysis of the H3K27ac modification level of COL1A2 promoter. (b) The mRNA expression of COL1A2 in parental cells by RT-qPCR. (c) The protein expression of COL1A2 in parental cells by western blot. (d) UCSC browser analysis of the binding of EP300 to the promoter of COL1A2. GC cells were transfected with oe-NC or oe-EP300. (e) ChIP-qPCR analysis of the H3K27ac modification level of COL1A2 promoter. (f) The mRNA expression of COL1A2 in parental cells by RT-qPCR. (g) The protein expression of COL1A2 in parental cells by western blot. (h) Colocalization of TWIST1 with EP300 in AGS and MKN-45 cells by double-labeled immunofluorescence staining. Data indicate means ± SD of three biological replicates. Two-way ANOVA and Tukey's multiple comparison test; ∗∗ p < 0.01 (vs. DMSO or oe-NC).

Article Snippet: Human GC cell lines MKN-45 and AGS were purchased from China Center for Type Culture Collection (Wuhan, Hubei, China).

Techniques: Expressing, Modification, Quantitative RT-PCR, Western Blot, Binding Assay, Transfection, Labeling, Immunofluorescence, Staining

Journal: Analytical Cellular Pathology (Amsterdam)

Article Title: TWIST1-EP300 Expedites Gastric Cancer Cell Resistance to Apatinib by Activating the Expression of COL1A2

doi: 10.1155/2022/5374262

Figure Lengend Snippet: Overexpression of COL1A2 promotes resistance of parental cells to apatinib. GC parental cells were transfected with oe-NC or oe-COL1A2. (a) The mRNA expression of COL1A2 in parental cells by RT-qPCR. (b) The protein expression of COL1A2 in parental cells by western blot. (c) The IC50 values of parental cells by CCK-8 assay. (d) The number of colonies formed by parental MKN-45 and AGS cells treated with 1 μ M apatinib. (e) Flow cytometry analysis of the proportion of apoptotic cells in parental MKN-45 and AGS cells. (f) TUNEL analysis of the number of apoptotic bodies in parental MKN-45 and AGS cells. Data indicate means ± SD of three biological replicates. Two-way ANOVA and Tukey's multiple comparison test; ∗∗ p < 0.01 (vs. oe-NC).

Article Snippet: Human GC cell lines MKN-45 and AGS were purchased from China Center for Type Culture Collection (Wuhan, Hubei, China).

Techniques: Over Expression, Transfection, Expressing, Quantitative RT-PCR, Western Blot, CCK-8 Assay, Flow Cytometry, TUNEL Assay

Journal: Analytical Cellular Pathology (Amsterdam)

Article Title: TWIST1-EP300 Expedites Gastric Cancer Cell Resistance to Apatinib by Activating the Expression of COL1A2

doi: 10.1155/2022/5374262

Figure Lengend Snippet: Knockdown of COL1A2 inhibits the sensitivity of drug-resistant cells to apatinib. GC-resistant cells were transfected with Scr or sh-COL1A2 #1 or #2. (a) The mRNA expression of COL1A2 in resistant cells by RT-qPCR. (b) The protein expression of COL1A2 in resistant cells by western blot. (c) The IC50 values of resistant cells by CCK-8 assay. (d) The number of colonies formed by resistant MKN-45 and AGS cells treated with 1 μ M apatinib. (e) Flow cytometry analysis of the proportion of apoptotic cells in resistant MKN-45 and AGS cells. (f) TUNEL analysis of the number of apoptotic bodies in resistant MKN-45 and AGS cells. Data indicate means ± SD of three biological replicates. Two-way ANOVA and Tukey's multiple comparison test; ∗∗ p < 0.01 (vs. Scr).

Article Snippet: Human GC cell lines MKN-45 and AGS were purchased from China Center for Type Culture Collection (Wuhan, Hubei, China).

Techniques: Transfection, Expressing, Quantitative RT-PCR, Western Blot, CCK-8 Assay, Flow Cytometry, TUNEL Assay

Journal: Analytical Cellular Pathology (Amsterdam)

Article Title: TWIST1-EP300 Expedites Gastric Cancer Cell Resistance to Apatinib by Activating the Expression of COL1A2

doi: 10.1155/2022/5374262

Figure Lengend Snippet: Knockdown of TWIST1 or EP300 reduces resistance to apatinib in parental cells overexpressing COL1A2. GC parental cells transfected with oe-COL1A2 were further transfected with sh-TWIST1 or sh-EP300. (a) The mRNA expression of TWIST1, EP300, and COL1A2 in parental cells by RT-qPCR. (b) The protein expression of TWIST1, EP300, and COL1A2 in parental cells by western blot. (c) The IC50 values of parental cells by CCK-8 assay. (d) The number of colonies formed by parental MKN-45 and AGS cells treated with 1 μ M apatinib. (e) Flow cytometry analysis of the proportion of apoptotic cells in parental MKN-45 and AGS cells. Data indicate means ± SD of three biological replicates. Two-way ANOVA and Tukey's multiple comparison test; ∗∗ p < 0.01 (vs. oe − COL1A2 + Scr).

Article Snippet: Human GC cell lines MKN-45 and AGS were purchased from China Center for Type Culture Collection (Wuhan, Hubei, China).

Techniques: Transfection, Expressing, Quantitative RT-PCR, Western Blot, CCK-8 Assay, Flow Cytometry

Journal: Bioengineered

Article Title: MiR-874-3p represses the migration and invasion yet promotes the apoptosis and cisplatin sensitivity via being sponged by long intergenic non-coding RNA 00922 (LINC00922) and targeting Glycerophosphodiester Phosphodiesterase Domain Containing 5 (GDPD5) in gastric cancer cells

doi: 10.1080/21655979.2022.2045831

Figure Lengend Snippet: GDPD5 was the target gene of miR-874-3p. (a) The downstream target gene of miR-874-3p was predicted and confirmed as well, and the databases, including TargetScan ( www.targetscan.org/vert_72 ), StarBase (starbase.sysu.edu.cn), and TCGA-STAD, www.genome.gov/Funded-Programs-Projects/Cancer-Genome-Atlas ) were utilized. The results concerning the possible downstream target genes were finally sorted and presented using a Venn diagram (A) . (b-c) The expression of the potential target genes of miR-874-3p after downregulating miR-874-3p was measured in DDP-resistant GC cell AGS (b) and HGC-27 (c) by qRT-PCR. GAPDH was used as the housekeeping gene. (d-f) Data from TargetScan (d) and dual-luciferase reporter assay (e-f) confirmed GDPD5 as the target gene of miR-874-3p. All data were expressed as mean ± standard deviation (SD), which was indicative of three independent tests. # p < 0.05, ## p < 0.01, ### p < 0.001, vs. IC; ^^^ p < 0.001, vs. MC. TCGA-STAD: The Cancer Genome Atlas Stomach Adenocarcinoma; USP43: ubiquitin specific peptidase 43; GDPD5: Glycerophosphodiester Phosphodiesterase Domain Containing 5; MYRF: myelin regulatory factor; PARP14: poly(ADP-ribose) polymerase family member 14; SATB2: SATB homeobox 2; RASGEF1A: RasGEF domain family member 1A; KCNC3: potassium voltage-gated channel subfamily C member 3; CDH11: cadherin 11.

Article Snippet: Human gastric mucosal epithelial cell line GES-1 (iCell-h062) as well as GC cell lines AGS (iCell-h016) and HGC-27 (iCell-h095) were bought from iCell (Shanghai, China, www.icellbioscience.com ).

Techniques: Expressing, Quantitative RT-PCR, Luciferase, Reporter Assay, Standard Deviation, Ubiquitin Proteomics

Journal: Bioengineered

Article Title: MiR-874-3p represses the migration and invasion yet promotes the apoptosis and cisplatin sensitivity via being sponged by long intergenic non-coding RNA 00922 (LINC00922) and targeting Glycerophosphodiester Phosphodiesterase Domain Containing 5 (GDPD5) in gastric cancer cells

doi: 10.1080/21655979.2022.2045831

Figure Lengend Snippet: MiR-874-3p was lower-expressed in drug-resistant GC, and miR-874-3p upregulation repressed the cell viability but enhanced sensitivity and apoptosis in GC cells with or without DDP. (a) Relative miR-874-3p expression in drug-resistant (Resistant) and drug-sensitive (Sensitive) GC tissues was quantified using qRT-PCR. U6 was employed as the internal control. (b) Relative miR-874-3p expression in DDP-resistant or parental GC cell and gastric mucosal epithelial cell line GES-1 was measured with qRT-PCR. U6 was applied as the internal control. (c-d) Relative miR-874-3p expression in DDP-resistant or parental GC cell AGS (c) and HGC-27 (d) after transfection was quantified with qRT-PCR. U6 was the internal control. (e-f) The effects of miR-874-3p on the viability of DDP-resistant or parental GC cell AGS (e) and HGC-27 (f) after the administration of DDP (0, 1.25, 2.5, 5, 10, 20, 40, 80 and 160 μmol/L) for 48 hours were assessed with CCK-8 assay. (g-h) The effects of miR-874-3p on the apoptosis of GC cell AGS (g) and HGC-27 (h) with or without 20 μmol/L DDP treatment for 48 hours were confirmed with flow cytometric assay. All data were expressed as mean ± standard deviation (SD), which was indicative of three independent tests. *** p < 0.001, vs. GES-1; ### p < 0.001, vs. AGS; &&& p < 0.001, vs. HGC-27; ^ p < 0.05, ^^ p < 0.01, ^^^ p < 0.001, vs. MC. miR: microRNA; GC: gastric cancer; DDP: cisplatin; qRT-PCR: quantitative real-time polymerase chain reaction; IC 50 : the half maximal inhibitory concentration; MC: mimic control; M: mimic; CCK-8: Cell Counting Kit-8.

Article Snippet: Human gastric mucosal epithelial cell line GES-1 (iCell-h062) as well as GC cell lines AGS (iCell-h016) and HGC-27 (iCell-h095) were bought from iCell (Shanghai, China, www.icellbioscience.com ).

Techniques: Expressing, Quantitative RT-PCR, Control, Transfection, CCK-8 Assay, Flow Cytometry, Standard Deviation, Real-time Polymerase Chain Reaction, Concentration Assay, Cell Counting

Journal: Bioengineered

Article Title: MiR-874-3p represses the migration and invasion yet promotes the apoptosis and cisplatin sensitivity via being sponged by long intergenic non-coding RNA 00922 (LINC00922) and targeting Glycerophosphodiester Phosphodiesterase Domain Containing 5 (GDPD5) in gastric cancer cells

doi: 10.1080/21655979.2022.2045831

Figure Lengend Snippet: MiR-874-3p upregulation suppressed the migration and invasion in GC cells with or without DDP. (a-b) The effects of miR-874-3p on the migration of DDP-resistant or parental GC cell AGS (a) and HGC-27 (b) were confirmed with Scratch assay at 0 and 48 hours under × 100 magnification (Scale bar = 50 μm). (c-d) The effects of miR-874-3p on the invasion of DDP-resistant or parental GC cell AGS (c) and HGC-27 (d) were unveiled with Transwell assay at 48 hours (× 250 magnification; Scale bar = 50 μm). All data were expressed as mean ± standard deviation (SD), which was indicative of three independent tests. ^^^ p < 0.001, vs. MC.

Article Snippet: Human gastric mucosal epithelial cell line GES-1 (iCell-h062) as well as GC cell lines AGS (iCell-h016) and HGC-27 (iCell-h095) were bought from iCell (Shanghai, China, www.icellbioscience.com ).

Techniques: Migration, Wound Healing Assay, Transwell Assay, Standard Deviation

Journal: Bioengineered

Article Title: MiR-874-3p represses the migration and invasion yet promotes the apoptosis and cisplatin sensitivity via being sponged by long intergenic non-coding RNA 00922 (LINC00922) and targeting Glycerophosphodiester Phosphodiesterase Domain Containing 5 (GDPD5) in gastric cancer cells

doi: 10.1080/21655979.2022.2045831

Figure Lengend Snippet: LINC00922 was the upstream lncRNA for miR-874-3p, and miR-874-3p downregulation eliminated the effects of LINC00922 silencing on miR-874-3p expression in DDP-resistant GC cells. (a-c) LncBase Predicted V2 ( http://carolina.imis.athena-innovation.gr/diana_tools ) (a) predicted and dual-luciferase reporter assay (b-c) confirmed LINC00922 as the upstream lncRNA for miR-874-3p. (d-g) Relative expressions of LINC00922 (d-e) and miR-874-3p (f-g) in DDP-resistant GC cell AGS and HGC-27 after transfection were quantified with qRT-PCR. GAPDH (for LINC00922) and U6 (for miR-874-3p) were the internal controls. (h-i) Relative miR-874-3p expression in DDP-resistant GC cell AGS (h) and HGC-27 (i) after transfection was quantified with qRT-PCR. (j-k) Relative LINC00922 expression in DDP-resistant GC cell AGS/DDP (j) and HGC-27/DDP (k) after transfection was quantified with qRT-PCR. U6 was used as the internal control. All data were expressed as mean ± standard deviation (SD), which was indicative of three independent tests. ^^ p < 0.01, ^^^ p < 0.001, vs. MC; *** p < 0.001, vs. siNC; ### p < 0.001, vs. IC; +++ p < 0.001, vs. siNC+IC; &&& p < 0.001, vs. siLINC00922 + I. LINC00922: long intergenic non-coding RNA 00922; LncRNA: long non-coding RNA; WT: wild-type; MUT: mutated; siRNA: small interfering RNA; IC: inhibitor control; I: inhibitor.

Article Snippet: Human gastric mucosal epithelial cell line GES-1 (iCell-h062) as well as GC cell lines AGS (iCell-h016) and HGC-27 (iCell-h095) were bought from iCell (Shanghai, China, www.icellbioscience.com ).

Techniques: Expressing, Luciferase, Reporter Assay, Transfection, Quantitative RT-PCR, Control, Standard Deviation, Small Interfering RNA

Journal: Bioengineered

Article Title: MiR-874-3p represses the migration and invasion yet promotes the apoptosis and cisplatin sensitivity via being sponged by long intergenic non-coding RNA 00922 (LINC00922) and targeting Glycerophosphodiester Phosphodiesterase Domain Containing 5 (GDPD5) in gastric cancer cells

doi: 10.1080/21655979.2022.2045831

Figure Lengend Snippet: MiR-874-3p downregulation eliminated the effects of LINC00922 silencing on cell viability and apoptosis in DDP-resistant GC cells (a-b) The effects of LINC00922 silencing and miR-874-3p downregulation on the viability in DDP-resistant GC cell AGS (a) and HGC-27 (b) after the administration of DDP (0, 1.25, 2.5, 5, 10, 20, 40, 80 and 160 μmol/L) for 48 hours were determined with CCK-8 assay. (c-d) The effects of LINC00922 silencing and miR-874-3p downregulation on the apoptosis of DDP-resistant GC cell AGS (c) and HGC-27 (d) were evaluated with flow cytometry. All data were expressed as mean ± standard deviation (SD), which was indicative of three independent tests. + p < 0.05, ++ p < 0.01, +++ p < 0.001, vs. siNC+IC; & p < 0.05, && p < 0.01, &&& p < 0.001, vs. siLINC00922 + I.

Article Snippet: Human gastric mucosal epithelial cell line GES-1 (iCell-h062) as well as GC cell lines AGS (iCell-h016) and HGC-27 (iCell-h095) were bought from iCell (Shanghai, China, www.icellbioscience.com ).

Techniques: CCK-8 Assay, Flow Cytometry, Standard Deviation

Journal: Bioengineered

Article Title: MiR-874-3p represses the migration and invasion yet promotes the apoptosis and cisplatin sensitivity via being sponged by long intergenic non-coding RNA 00922 (LINC00922) and targeting Glycerophosphodiester Phosphodiesterase Domain Containing 5 (GDPD5) in gastric cancer cells

doi: 10.1080/21655979.2022.2045831

Figure Lengend Snippet: Downregulated miR-874-3p eliminated the effects of LINC00922 silencing on cell migration and invasion in DDP-resistant GC cells. (a-b) The effects of LINC00922 silencing and miR-874-3p downregulation on the migration of DDP-resistant GC cell AGS (a) and HGC-27 (b) were confirmed with Scratch assay at 0 and 48 hours, under × 100 magnification (Scale bar = 50 μm). (c-d) The effects of LINC00922 silencing and miR-874-3p downregulation on the invasion of DDP-resistant GC cell AGS (c) and HGC-27 (d) at 48 hours were detected with Transwell assay (× 250 magnification; Scale bar = 50 μm). All data were expressed as mean ± standard deviation (SD), which was indicative of three independent tests. +++ p < 0.001, vs. siNC+IC; &&& p < 0.001, vs. siLINC00922 + I.

Article Snippet: Human gastric mucosal epithelial cell line GES-1 (iCell-h062) as well as GC cell lines AGS (iCell-h016) and HGC-27 (iCell-h095) were bought from iCell (Shanghai, China, www.icellbioscience.com ).

Techniques: Migration, Wound Healing Assay, Transwell Assay, Standard Deviation

Journal: Bioengineered

Article Title: MiR-874-3p represses the migration and invasion yet promotes the apoptosis and cisplatin sensitivity via being sponged by long intergenic non-coding RNA 00922 (LINC00922) and targeting Glycerophosphodiester Phosphodiesterase Domain Containing 5 (GDPD5) in gastric cancer cells

doi: 10.1080/21655979.2022.2045831

Figure Lengend Snippet: MiR-874-3p downregulation eliminated the effects of LINC00922 silencing on GDPD5 in DDP-resistant GC cells, the expression of which was also decreased via transfection. (a-d) The effects of LINC00922 silencing and miR-874-3p downregulation on GDPD5 expression of DDP-resistant GC cell AGS/DDP (a-b) and HGC-27/DDP (c-d) were determined by qRT-PCR and Western blot. GAPDH was the housekeeping gene. (e-h) GDPD5 expression was measured again in DDP-resistant GC cell AGS/DDP and HGC-27/DDP following the transfection of siGDPD5, as evidenced by qRT-PCR (e-f) and Western blot (g-h). GAPDH was the housekeeping gene. All data were expressed as mean ± standard deviation (SD), which was indicative of three independent tests. +++ p < 0.001, vs. siNC+IC; &&& p < 0.001, vs. siLINC00922 + I; ^^^ p < 0.001, vs. siNC.

Article Snippet: Human gastric mucosal epithelial cell line GES-1 (iCell-h062) as well as GC cell lines AGS (iCell-h016) and HGC-27 (iCell-h095) were bought from iCell (Shanghai, China, www.icellbioscience.com ).

Techniques: Expressing, Transfection, Quantitative RT-PCR, Western Blot, Standard Deviation

Journal: Bioengineered

Article Title: MiR-874-3p represses the migration and invasion yet promotes the apoptosis and cisplatin sensitivity via being sponged by long intergenic non-coding RNA 00922 (LINC00922) and targeting Glycerophosphodiester Phosphodiesterase Domain Containing 5 (GDPD5) in gastric cancer cells

doi: 10.1080/21655979.2022.2045831

Figure Lengend Snippet: GDPD5 silencing diminished the effects of downregulated miR-874-3p on GDPD5 expression and cell viability of DDP-resistant GC cells. (a-d) The effects of GDPD5 silencing and miR-874-3p downregulation on GDPD5 expression in DDP-resistant GC cell AGS/DDP (a-b) and HGC-27/DDP (c-d) were unveiled as confirmation of qRT-PCR and Western blot. GAPDH was used as the internal control. (e-f) The effects of GDPD5 silencing and miR-874-3p downregulation on the cell viability in DDP-resistant GC cell AGS/DDP (e) and HGC-27/DDP (f) were suggested by CCK-8 assay. All data were expressed as mean ± standard deviation (SD), which was indicative of three independent tests. + p < 0.05, ++ p < 0.01, +++ p < 0.001, vs. siNC+IC; # p < 0.05, ## p < 0.01, ### p < 0.001, vs. siGDPD5 + I.

Article Snippet: Human gastric mucosal epithelial cell line GES-1 (iCell-h062) as well as GC cell lines AGS (iCell-h016) and HGC-27 (iCell-h095) were bought from iCell (Shanghai, China, www.icellbioscience.com ).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Control, CCK-8 Assay, Standard Deviation

Journal: Bioengineered

Article Title: MiR-874-3p represses the migration and invasion yet promotes the apoptosis and cisplatin sensitivity via being sponged by long intergenic non-coding RNA 00922 (LINC00922) and targeting Glycerophosphodiester Phosphodiesterase Domain Containing 5 (GDPD5) in gastric cancer cells

doi: 10.1080/21655979.2022.2045831

Figure Lengend Snippet: GDPD5 silencing diminished the effects of downregulated miR-874-3p on the migration and invasion of DDP-resistant GC cells. (a-b) The effects of GDPD5 silencing and miR-874-3p downregulation on the migration of DDP-resistant GC cells AGS/DDP (a) and HGC-27/DDP (b) were confirmed with Scratch assay at 0 and 48 hours, under × 100 magnification (Scale bar = 100 μm). (c-d) The effects of GDPD5 silencing and miR-874-3p downregulation on the invasion of DDP-resistant GC cells AGS (c) and HGC-27 (d) at 48 hours were detected with Transwell assay (× 250 magnification; Scale bar = 50 μm). All data were expressed as mean ± standard deviation (SD), which was indicative of three independent tests. + p < 0.05, ++ p < 0.01, +++ p < 0.001, vs. IC+siNC; & p < 0.05, && p < 0.01, &&& p < 0.001, vs. I+ siGDPD5.

Article Snippet: Human gastric mucosal epithelial cell line GES-1 (iCell-h062) as well as GC cell lines AGS (iCell-h016) and HGC-27 (iCell-h095) were bought from iCell (Shanghai, China, www.icellbioscience.com ).

Techniques: Migration, Wound Healing Assay, Transwell Assay, Standard Deviation

Journal: Engineering in Life Sciences

Article Title: Overexpression of GRP78 enhances survival of CHO cells in response to serum deprivation and oxidative stress

doi: 10.1002/elsc.201500152

Figure Lengend Snippet: Viability of Chinese hamster ovary (CHO) cells in response to serum deprivation and oxidative stress. CHO cells were seeded in 12‐well plates at a density of 1.5 × 105 cells per well and grown for 24 h. Then cells were cultured in Roswell Park Memorial Institute (RPMI) 1640 medium, serum free medium with or without 200 μM H2O2. In order to ensure that other nutrient limitations were not experienced, cells were fed fresh medium every 48 h. Cells were taken every 2 days and analyzed for cell viability using Annexin V/PI double staining. Error bars represent SD for each measurement.

Article Snippet: The Chinese hamster ovary (CHO) cells, human stomach adenocarcinoma (AGS) cell line, and human hepatoma cell line HepG2 (China Center for Type Culture Collection, Wuhan, P. R. China) were cultured in Roswell Park Memorial Institute (RPMI) 1640 and DMEM medium (Gbico BRL, NY, USA) supplemented with 10% FBS (Sijiqing, Hangzhou, China) and 100 U/mL ampicillin, 100 mg/mL streptomycin in an incubator containing 5% CO 2 at 37°C (standard conditions).

Techniques: Cell Culture, Double Staining

Journal: Engineering in Life Sciences

Article Title: Overexpression of GRP78 enhances survival of CHO cells in response to serum deprivation and oxidative stress

doi: 10.1002/elsc.201500152

Figure Lengend Snippet: Glucose‐regulated protein 78 (GRP78) engineering protected Chinese hamster ovary (CHO) cells against injury induced by H2O2. (A) Cells synchronized in serum‐free medium were treated with H2O2 at increasing concentration for 4 h and then cultured with 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide (MTT; 5 mg/mL) for 4 h. Bar graph represented the percentage of viable cells assayed by MTT. (B) The level of cleaved caspase‐3 in H2O2‐treated cells was detected by western blot (upper). The lower left bar graph represented densitometric analysis of GRP78 levels of the western blots and the lower right of cleaved caspase‐3. Data were mean values ± SD, n > 3.*p < 0.05,**p < 0.01,***p < 0.001, NS, nonsignificant difference versus negative control.

Article Snippet: The Chinese hamster ovary (CHO) cells, human stomach adenocarcinoma (AGS) cell line, and human hepatoma cell line HepG2 (China Center for Type Culture Collection, Wuhan, P. R. China) were cultured in Roswell Park Memorial Institute (RPMI) 1640 and DMEM medium (Gbico BRL, NY, USA) supplemented with 10% FBS (Sijiqing, Hangzhou, China) and 100 U/mL ampicillin, 100 mg/mL streptomycin in an incubator containing 5% CO 2 at 37°C (standard conditions).

Techniques: Concentration Assay, Cell Culture, Western Blot, Negative Control

Journal: Engineering in Life Sciences

Article Title: Overexpression of GRP78 enhances survival of CHO cells in response to serum deprivation and oxidative stress

doi: 10.1002/elsc.201500152

Figure Lengend Snippet: Glucose‐regulated protein 78 (GRP78) engineering enhanced yields of TfR‐Ab and improved cell viability. Chinese hamster ovary (CHO) and CHO modified by GRP78 (CHO‐GRP78) cells were seeded in 12‐well plates at a density of 2 × 105 cells per well and transiently transfected with pOptiVEC™‐TOPO®/TfR‐Ab, followed by culturing with SFM4CHO™ medium. (A) The concentration of TfR‐Ab in the supernatant was detected by ELISA assay. (B) The quality of TfR‐Ab in the supernatant was evaluated by its binding ability with TfR+ HepG2 cells using flow cytometry (FCM). The bar graphs represented FCM analysis of mean fluorescence index (MFI). (C) Bar graph represented FCM analysis of the percentage viability of CHO and CHO‐GRP78 cells. (D) The viable cell numbers of CHO and CHO‐GRP78 cells were counted after Trypan blue staining. Data were mean values ± SD, n > 3.*p < 0.05,**p < 0.01,***p < 0.001 versus negative control.

Article Snippet: The Chinese hamster ovary (CHO) cells, human stomach adenocarcinoma (AGS) cell line, and human hepatoma cell line HepG2 (China Center for Type Culture Collection, Wuhan, P. R. China) were cultured in Roswell Park Memorial Institute (RPMI) 1640 and DMEM medium (Gbico BRL, NY, USA) supplemented with 10% FBS (Sijiqing, Hangzhou, China) and 100 U/mL ampicillin, 100 mg/mL streptomycin in an incubator containing 5% CO 2 at 37°C (standard conditions).

Techniques: Modification, Transfection, Concentration Assay, Enzyme-linked Immunosorbent Assay, Binding Assay, Flow Cytometry, Fluorescence, Staining, Negative Control