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Cellular expression and functional validation of CTHRC1 in vitro . (A–C) Quantitative reverse-transcription polymerase chain reaction (qRT-PCR) analysis of SERPINE1, CTHRC1, and EGR1 in GES-1, <t>AGS,</t> HGC-27, and MKN-45 cells. (D) qRT-PCR screening of three CTHRC1-targeting short hairpin RNAs (shRNAs) in HGC-27 and <t>AGS</t> <t>cells.</t> (E) Western blot validation of CTHRC1 knockdown; glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was the loading control. (F, G) Cell Counting Kit-8 (CCK-8) proliferation assays. (H, I) Representative 5-ethynyl-2′-deoxyuridine (EdU) staining and quantification; scale bar, 100 µm. (J, K) Colony-formation assays and quantification; scale bar, 10 mm. (L–N) Wound-healing assays and migration-rate quantification; scale bar, 100 µm. (O–R) Transwell migration/invasion assays and quantification; scale bar, 100 µm. Data are mean ± standard deviation (SD) from three independent biological experiments. (A–D) were analyzed by one-way analysis of variance (ANOVA) with Dunnett correction, panels F and G by two-way ANOVA with multiple-comparison correction, and two-group endpoint comparisons by two-sided Student’s t-tests. ***P < 0.001.
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Cellular expression and functional validation of CTHRC1 in vitro . (A–C) Quantitative reverse-transcription polymerase chain reaction (qRT-PCR) analysis of SERPINE1, CTHRC1, and EGR1 in GES-1, AGS, HGC-27, and MKN-45 cells. (D) qRT-PCR screening of three CTHRC1-targeting short hairpin RNAs (shRNAs) in HGC-27 and AGS cells. (E) Western blot validation of CTHRC1 knockdown; glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was the loading control. (F, G) Cell Counting Kit-8 (CCK-8) proliferation assays. (H, I) Representative 5-ethynyl-2′-deoxyuridine (EdU) staining and quantification; scale bar, 100 µm. (J, K) Colony-formation assays and quantification; scale bar, 10 mm. (L–N) Wound-healing assays and migration-rate quantification; scale bar, 100 µm. (O–R) Transwell migration/invasion assays and quantification; scale bar, 100 µm. Data are mean ± standard deviation (SD) from three independent biological experiments. (A–D) were analyzed by one-way analysis of variance (ANOVA) with Dunnett correction, panels F and G by two-way ANOVA with multiple-comparison correction, and two-group endpoint comparisons by two-sided Student’s t-tests. ***P < 0.001.

Journal: Frontiers in Immunology

Article Title: A palmitoylation-associated three-gene signature predicts prognosis, immune features, and drug sensitivity in gastric cancer: functional validation of CTHRC1

doi: 10.3389/fimmu.2026.1893438

Figure Lengend Snippet: Cellular expression and functional validation of CTHRC1 in vitro . (A–C) Quantitative reverse-transcription polymerase chain reaction (qRT-PCR) analysis of SERPINE1, CTHRC1, and EGR1 in GES-1, AGS, HGC-27, and MKN-45 cells. (D) qRT-PCR screening of three CTHRC1-targeting short hairpin RNAs (shRNAs) in HGC-27 and AGS cells. (E) Western blot validation of CTHRC1 knockdown; glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was the loading control. (F, G) Cell Counting Kit-8 (CCK-8) proliferation assays. (H, I) Representative 5-ethynyl-2′-deoxyuridine (EdU) staining and quantification; scale bar, 100 µm. (J, K) Colony-formation assays and quantification; scale bar, 10 mm. (L–N) Wound-healing assays and migration-rate quantification; scale bar, 100 µm. (O–R) Transwell migration/invasion assays and quantification; scale bar, 100 µm. Data are mean ± standard deviation (SD) from three independent biological experiments. (A–D) were analyzed by one-way analysis of variance (ANOVA) with Dunnett correction, panels F and G by two-way ANOVA with multiple-comparison correction, and two-group endpoint comparisons by two-sided Student’s t-tests. ***P < 0.001.

Article Snippet: AGS cells were cultured in F-12K medium (ATCC, 30-2004) supplemented with 10% fetal bovine serum (FBS; ATCC, 30-2020) and 1% penicillin-streptomycin (Gibco, 15140122).

Techniques: Expressing, Functional Assay, Biomarker Discovery, In Vitro, Reverse Transcription, Polymerase Chain Reaction, Quantitative RT-PCR, Analysis, Screening, Western Blot, Knockdown, Control, Cell Counting, CCK-8 Assay, Staining, Colony Assay, Migration, Standard Deviation, Comparison