human epithelial gastric cells ags  (ATCC)

Journal: Biochemical and biophysical research communications

Article Title: Helicobacter pylori virulence factor CagA promotes Snail-mediated epithelial-mesenchymal transition and invasive behavior by downregulating Semaphorin 5A in gastric epithelial cells.

doi: 10.1016/j.bbrc.2025.151421

Figure Lengend Snippet: Fig. 3. Infection with CagA-positive H. pylori as well as knockout (KO) of the SEMA5A gene induces expression of Snail and N-cadherin to promote invasive behavior in gastric epithelial cells. (A) Western blot analysis of expression of SEMA5A, EMT-driving transcription factor Snail, mesenchymal marker N-cadherin in WT TN2- infected and ΔCagA TN2-infected gastric epithelial cells. GAPDH was used as a loading control. (B) Giemsa staining of WT TN2-infected and ΔCagA TN2-infected gastric epithelial cells that invaded to the lower surface of the membrane (upper panel). Fold change of the number of cells shown to invade to the lower surface of the membrane is demonstrated. Error bars represent mean SD (n = 3) and P values were determined by Student’s t-test (lower panel). (C) Knockout (KO) of the SEMA5A gene in AGS gastric epithelial cells. Structure of the SEMA5A gene, guide RNA (gRNA) targeting sequence and PAM sequence in exon 6 are indicated (upper panel). Sanger sequencing of the gRNA targeting site in intact and SEMA5A KO cells. SEMA5A KO cells possess a biallelic 1-bp deletion in exon 6 (lower left panel). Real-time quantitative RT-PCR analysis of expression of SEMA5A in intact and SEMA5A KO cells (lower middle panel). GAPDH was used as endogenous control to calculate relative expression of SEMA5A. Error bars represent mean SD (n = 3) and P values were determined by Student’s t-test. Western blot analysis of expression of SEMA5A. GAPDH was used as a loading control (lower right panel). (D) Western blot analysis of expression of Snail and N-cadherin in intact and SEMA5A KO gastric epithelial cells. GAPDH was used as a loading control. (E) Giemsa staining of intact and SEMA5A KO gastric epithelial cells that invaded to the lower surface of the membrane (upper panel). Fold change of the number of cells shown to invade to the lower surface of the membrane is demonstrated. Error bars represent mean SD (n = 3) and P values were determined by Student’s t-test (lower panel).

Article Snippet: Human gastric epithelial cell lines AGS (ATCC CRL1739) and AGS SEMA5A KO were maintained in RPMI 1640 with 10 % fetal bovine serum (FBS) at 37◦C in humidified atmosphere with 5 % CO2.

Techniques: Infection, Knock-Out, Expressing, Western Blot, Marker, Control, Staining, Membrane, Sequencing, Quantitative RT-PCR

Journal: Frontiers in Pharmacology

Article Title: Plukenetia volubilis leaves as source of anti- Helicobacter pylori agents

doi: 10.3389/fphar.2024.1461447

Figure Lengend Snippet: P. volubilis water infusion, EA extract and astragalin do not affect the viability of AGS human epithelial cells. AGS cells were treated with increasing concentrations of P. volubilis water infusion (A) , ethyl acetate extract (B) and astragalin (C) and analysed following MTT assay. Results are presented as mean and standard deviation (error bars) of triplicate values and were compared using two-way ANOVA with the Sidak’s multiple comparison test. No statistically significant differences were observed.

Article Snippet: AGS human epithelial gastric adenocarcinoma cells (ATCC CRL-1739), with a passage number <20, were maintained in Ham’s F12 nutrient mix (Sigma) supplemented with 10% FBS and cultured in a 5% CO2 humidified 37°C incubator.

Techniques: MTT Assay, Standard Deviation, Comparison