Journal: bioRxiv

Article Title: Evaluating Spike Antigenicity across Endemic Human Coronavirus Models using Flow Virometry

doi: 10.64898/2026.05.28.728498

Figure Lengend Snippet: (A) Supernatants from HCoV-229E-infected human cell lines (MRC-5, Huh7.5, and HCT-8; top to bottom rows) were stained using S-specific mAbs targeting the RBD (C04), S1 (F07), or S2 (F12) domains, or an isotype control (IgG), followed by a PE-labeled secondary antibody. Gates for positive staining were set above the isotype control background (>15 PE MESF on the y-axis). Calibrated mean PE fluorescence values (MESF) from gated regions are shown in red. (B) Quantitative comparison of staining results from the gates in (A), with bars representing mean PE MESF ± standard deviation wherein light blue = MRC-5, blue = Huh7.5, and dark blue = HCT-8. Data are representative of at least three independent replicates. Statistical significance was determined using two-way ANOVA with Dunnett’s multiple comparisons test to compare each antibody condition to the IgG control within each virus. Significance thresholds: **p < 0.01, ***p < 0.001, ****p < 0.0001.(C) HCoV-229E pseudoviruses produced by transfection of 293T cells were stained directly in cell culture supernatants using the same antibody panel and staining protocol as in (A). (D) Quantification of staining in (C), with bars indicating mean PE MESF ± standard deviation. See also Figures S1 - S3.

Article Snippet: HCT-8 (HRT-18) cells (ATCC, Cat# CCL-244) were cultured in RPMI (Gibco, Cat#11875-093).

Techniques: Infection, Staining, Control, Labeling, Fluorescence, Comparison, Standard Deviation, Virus, Produced, Transfection, Cell Culture

Journal: International Journal of Molecular Sciences

Article Title: SLC25A5 Suppresses Colorectal Cancer Growth and Metastasis Through Regulation of the EIF3A/PI3K/AKT Axis

doi: 10.3390/ijms27104334

Figure Lengend Snippet: SLC25A5 inhibits CRC progression by suppressing proliferation and promoting apoptosis: ( A , B ) Western blot ( A ) and RT-qPCR ( B ) analysis confirming the successful overexpression of SLC25A5 in HCT-8 and HT-29 cell lines. ( C , D ) CCK-8 proliferation assays showing the growth rates of HCT-8 ( C ) and HT-29 ( D ) cells transfected with either empty vector or SLC25A5 expression plasmid. ( E – H ) Representative images ( E , F ) and quantitative analysis ( G , H ) of EdU incorporation assays in HCT-8 and HT-29 cells (Nuclei were stained with DAPI (blue), and proliferating cells were stained with EdU (red). Scale bars: 200 μm). ( I ) Representative images and statistical analysis of colony formation assays. ( J ) Flow cytometry analysis and quantification of apoptotic cells in the indicated groups. ( K ) Western blot analysis of Bcl-2, Bax, and Cleaved-caspase3 (C-caspase3) protein levels in HCT-8 and HT-29 cells. ( L – N ) Representative images of xenograft tumors ( L ), tumor growth curves ( M ), and final tumor weights ( N ) from nude mice injected with the indicated HCT-8 cells (n = 6 per group). ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Human CRC cell lines HCT-8 and HT-29 were obtained from ATCC and cultured in RPMI-1640 or McCoy’s 5A medium supplemented with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin at 37 °C in a humidified incubator containing 5% CO 2 .

Techniques: Western Blot, Quantitative RT-PCR, Over Expression, CCK-8 Assay, Transfection, Plasmid Preparation, Expressing, Staining, Flow Cytometry, Injection

Journal: International Journal of Molecular Sciences

Article Title: SLC25A5 Suppresses Colorectal Cancer Growth and Metastasis Through Regulation of the EIF3A/PI3K/AKT Axis

doi: 10.3390/ijms27104334

Figure Lengend Snippet: SLC25A5 inhibits CRC metastasis by suppressing EMT and cell motility: ( A , B ) Representative images ( A ) and statistical analysis ( B ) of wound healing assays in HCT-8 and HT-29 cells at 0, 24, and 48 h (Scale bars: 500 μm). ( C , D ) Representative images ( C ) and quantitative analysis ( D ) of Transwell migration and Matrigel invasion assays (Scale bars: 100 μm). ( E ) Immunofluorescence staining of E-cadherin (orange) and N-cadherin (red) in the indicated CRC cells; nuclei were stained with DAPI (blue) (Scale bars: 50 μm). ( F ) Western blot analysis of E-cadherin, N-cadherin, and Vimentin protein levels in HCT-8 and HT-29 cells. ( G ) Representative images of livers and H&E-stained liver sections from the in vivo metastasis model. ( H , I ) Quantification of the number of metastatic nodules per area ( H ) and the percentage of tumor/liver area ( I ). * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Human CRC cell lines HCT-8 and HT-29 were obtained from ATCC and cultured in RPMI-1640 or McCoy’s 5A medium supplemented with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin at 37 °C in a humidified incubator containing 5% CO 2 .

Techniques: Migration, Immunofluorescence, Staining, Western Blot, In Vivo

Journal: International Journal of Molecular Sciences

Article Title: SLC25A5 Suppresses Colorectal Cancer Growth and Metastasis Through Regulation of the EIF3A/PI3K/AKT Axis

doi: 10.3390/ijms27104334

Figure Lengend Snippet: SLC25A5 Modulates the Proteasomal Degradation of EIF3A: ( A ) Silver staining showing protein bands pulled down by SLC25A5 in HCT-8 cells; arrows indicate EIF3A and SLC25A5. ( B ) Endogenous Co-IP analysis demonstrating the interaction between SLC25A5 and EIF3A in CRC cells. ( C ) Immunofluorescence images showing the co-localization of EIF3A (red) and SLC25A5 (green) in HCT-8 and HT-29 cells (Nuclei were counterstained with DAPI (blue). The yellow color in the merged images indicates the co-localization of EIF3A and SLC25A5 proteins. Scale bars: 10 μm). ( D , E ) RT-qPCR ( D ) and Western blot ( E ) analysis of EIF3A expression in CRC cells overexpressing SLC25A5. ( F ) Western blot analysis of EIF3A in SLC25A5-overexpressing cells treated with MG132 (10 μM) or CQ (50 μM). ( G , H ) Representative Western blot ( G ) and quantitative degradation curves ( H ) of EIF3A in HCT-8 cells treated with CHX (100 μg/mL) over a 24-h time course. ( I ) Ubiquitination assay showing the effect of SLC25A5 overexpression on the poly-ubiquitination of EIF3A in HCT-8 cells. ( J , K ) Representative Western blot ( J ) and quantitative degradation curves ( K ) of EIF3A in HT-29 cells treated with CHX (100 μg/mL). ( L ) Ubiquitination assay showing SLC25A5-mediated EIF3A poly-ubiquitination in HT-29 cells. ( M ) Representative IHC images showing the expression of SLC25A5 and EIF3A in two clinical CRC cases (Scale bars: 100 μm). ( N ) Pearson correlation analysis between SLC25A5 and EIF3A IHC scores in CRC specimens (n = 30). *** p < 0.001, ns, non-significant.

Article Snippet: Human CRC cell lines HCT-8 and HT-29 were obtained from ATCC and cultured in RPMI-1640 or McCoy’s 5A medium supplemented with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin at 37 °C in a humidified incubator containing 5% CO 2 .

Techniques: Silver Staining, Co-Immunoprecipitation Assay, Immunofluorescence, Quantitative RT-PCR, Western Blot, Expressing, Ubiquitin Proteomics, Over Expression

Journal: International Journal of Molecular Sciences

Article Title: SLC25A5 Suppresses Colorectal Cancer Growth and Metastasis Through Regulation of the EIF3A/PI3K/AKT Axis

doi: 10.3390/ijms27104334

Figure Lengend Snippet: SLC25A5 Suppresses CRC Progression in Association with EIF3A: ( A – D ) Representative images ( A , C ) and quantitative analysis ( B , D ) of EdU incorporation assays in HCT-8 and HT-29 cells transfected with the indicated plasmids (Nuclei were stained with DAPI (blue), and proliferating cells were stained with EdU (red). Scale bars: 200 μm). ( E , F ) Colony formation assays of CRC cells after the indicated treatments. ( G ) Western blot analysis of Bcl-2, Bax, and cleaved-caspase3 protein levels in rescue experiments. ( H – J ) Representative images of xenograft tumors ( H ), tumor growth curves ( I ), and final tumor weights ( J ) from the indicated groups. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Human CRC cell lines HCT-8 and HT-29 were obtained from ATCC and cultured in RPMI-1640 or McCoy’s 5A medium supplemented with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin at 37 °C in a humidified incubator containing 5% CO 2 .

Techniques: Transfection, Staining, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: SLC25A5 Suppresses Colorectal Cancer Growth and Metastasis Through Regulation of the EIF3A/PI3K/AKT Axis

doi: 10.3390/ijms27104334

Figure Lengend Snippet: SLC25A5 Suppresses CRC Metastasis and EMT in Association with EIF3A: ( A – C ) Representative images ( A ) and quantification ( B , C ) of Transwell invasion assays in CRC cells after the indicated treatments (Scale bars: 100 μm). ( D – F ) Representative images ( D ) and quantitative analysis ( E , F ) of wound healing assays. ( G , H ) Immunofluorescence staining of E-cadherin (orange) and N-cadherin (red) in HCT-8 ( G ) and HT-29 ( H ) cells (Nuclei were counterstained with DAPI (blue). Scale bars: 50 μm). ( I ) Western blot analysis of EIF3A, E-cadherin, N-cadherin, and Vimentin protein levels in rescue groups. ( J ) Representative images of liver metastases and H&E staining from the indicated groups. ( K ) Statistical analysis of the number of metastatic nodules and tumor/liver area ratio. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Human CRC cell lines HCT-8 and HT-29 were obtained from ATCC and cultured in RPMI-1640 or McCoy’s 5A medium supplemented with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin at 37 °C in a humidified incubator containing 5% CO 2 .

Techniques: Immunofluorescence, Staining, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: SLC25A5 Suppresses Colorectal Cancer Growth and Metastasis Through Regulation of the EIF3A/PI3K/AKT Axis

doi: 10.3390/ijms27104334

Figure Lengend Snippet: SLC25A5 Suppresses CRC Progression in Association with the EIF3A–PI3K–AKT Axis: ( A ) Volcano plot identifying differentially expressed genes (DEGs) after SLC25A5 overexpression in HCT-8 cells. ( B ) Top 10 enriched KEGG pathways based on the DEGs. ( C ) Immunofluorescence staining of E-cadherin and N-cadherin in cells treated with the AKT activator SC79 (Nuclei were counterstained with DAPI (blue). E-cadherin and N-cadherin were labeled with orange and red fluorescence, respectively. Scale bars: 50 μm). ( D ) Western blot analysis of SLC25A5, EIF3A, P-AKT, AKT, and EMT markers in the indicated groups. ( E ) Transwell invasion assays of CRC cells treated with DMSO or SC79 (Scale bars: 100 μm). ( F – H ) Representative liver images ( F ) and quantification of liver metastatic nodules ( G ) and tumor/liver area ratio ( H ) in the indicated groups. ** p < 0.01, **** p < 0.0001.

Article Snippet: Human CRC cell lines HCT-8 and HT-29 were obtained from ATCC and cultured in RPMI-1640 or McCoy’s 5A medium supplemented with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin at 37 °C in a humidified incubator containing 5% CO 2 .

Techniques: Over Expression, Immunofluorescence, Staining, Labeling, Fluorescence, Western Blot