Journal: Biology Direct

Article Title: Single-cell RNA sequencing reveals that TAF7 promotes colorectal cancer stemness by targeting SOX2

doi: 10.1186/s13062-026-00910-4

Figure Lengend Snippet: Primer sequences

Article Snippet: Following centrifugation, the supernatant was subjected to an overnight incubation with primary antibodies [rabbit anti-TAF7 (Cat# 13506-1-AP, Proteintech, USA) or rabbit anti-IgG (Cat# 10284-1-AP, Proteintech, USA)] at 4 °C with gentle rotation.

Techniques:

Journal: Biology Direct

Article Title: Single-cell RNA sequencing reveals that TAF7 promotes colorectal cancer stemness by targeting SOX2

doi: 10.1186/s13062-026-00910-4

Figure Lengend Snippet: Differential gene analysis of epithelial cell subpopulations in single-cell sequencing. A : UMAP plot of epithelial sub-clustering; B : UMAP plot of different sample groups for epithelial cells; C : UMAP plot of annotated epithelial subpopulations; D : Marker genes for epithelial subpopulations; E : Bar plot showing epithelial subpopulation proportions in different sample groups; F : Expression of differential genes in the TAF7 + EPC subpopulation; G : Malignancy score analysis of different epithelial subpopulations using the R package “infercnv”. TAF7 + EPC, TAF7-positive tumor epithelial cells

Article Snippet: Following centrifugation, the supernatant was subjected to an overnight incubation with primary antibodies [rabbit anti-TAF7 (Cat# 13506-1-AP, Proteintech, USA) or rabbit anti-IgG (Cat# 10284-1-AP, Proteintech, USA)] at 4 °C with gentle rotation.

Techniques: Analysis, Single-Cell Sequencing, Marker, Expressing

Journal: Biology Direct

Article Title: Single-cell RNA sequencing reveals that TAF7 promotes colorectal cancer stemness by targeting SOX2

doi: 10.1186/s13062-026-00910-4

Figure Lengend Snippet: TAF7 knockdown inhibits CRC cell stemness. A : RT-qPCR detection of TAF7 expression in HIEC-6, SW480, LoVo, and HT-29 cells; n = 3 independent experiments. Cell groups: sh-NC, sh-TAF7; B-C : RT-qPCR ( B ) and western blotting ( C ) confirming TAF7 knockdown; n = 3 independent experiments. D-E : RT-qPCR ( D ) and western blotting ( E ) detection of SOX2 mRNA and protein levels; n = 3 independent experiments. F : Sphere formation assay assessing in vitro sphere-forming ability, a total of 4 × 10⁴ cells were seeded per well, followed by incubation for 10 to 14 days; n = 3 independent experiments. G : Flow cytometry detection of stemness marker CD133 expression; n = 3 independent experiments. H : Transwell assay assessing cell migration and invasion (1 × 10⁵ cells were seeded into the upper chamber, and cell counting was performed 24 h later); n = 3 independent experiments. I : Wound healing assay assessing cell migration; n = 3 independent experiments. J : Western blotting detection of CD44, NANOG, E-cadherin and N-cadherin protein levels; n = 3 independent experiments. All quantitative data are presented as mean ± SD. Student’s t -test was used for comparisons between two groups, *, P < 0.05

Article Snippet: Following centrifugation, the supernatant was subjected to an overnight incubation with primary antibodies [rabbit anti-TAF7 (Cat# 13506-1-AP, Proteintech, USA) or rabbit anti-IgG (Cat# 10284-1-AP, Proteintech, USA)] at 4 °C with gentle rotation.

Techniques: Knockdown, Quantitative RT-PCR, Expressing, Western Blot, Tube Formation Assay, In Vitro, Incubation, Flow Cytometry, Marker, Transwell Assay, Migration, Cell Counting, Wound Healing Assay

Journal: Biology Direct

Article Title: Single-cell RNA sequencing reveals that TAF7 promotes colorectal cancer stemness by targeting SOX2

doi: 10.1186/s13062-026-00910-4

Figure Lengend Snippet: TAF7 promotes CRC cell stemness by transcriptionally activating SOX2. A : Binding sites between TAF7 and SOX2. B : ChIP assay showing TAF7 enrichment on the SOX2 promoter region, IgG served as negative control; n = 3 independent experiments. C : Dual-luciferase reporter assay determining interaction between TAF7 and SOX2; n = 3 independent experiments. Cell groups: oe-NC, oe-TAF7, oe-TAF7 + oe-SOX2-WT, oe-TAF7 + oe-SOX2-MUT; D : RT-qPCR detection of SOX2 mRNA expression; Cell groups: oe-NC + sh-NC, oe-TAF7 + sh-NC, oe-NC + sh-SOX2, oe-TAF7 + sh-SOX2; E : RT-qPCR detection of SOX2 mRNA expression; n = 3 independent experiments. F : Western blotting detection of CD44 and NANOG protein levels; n = 3 independent experiments. G : Sphere formation assay, a total of 4 × 10⁴ cells were seeded per well, followed by incubation for 10 to 14 days; n = 3 independent experiments. H : Flow cytometry detection of CD133 expression; n = 3 independent experiments. I : Transwell assay assessing migration and invasion (1 × 10⁵ cells were seeded into the upper chamber, and cell counting was performed 24 h later); = 3 independent experiments. J : Wound healing assay assessing migration; n = 3 independent experiments. K : Western blotting detection of E-cadherin and N-cadherin protein levels; n = 3 independent experiments. All quantitative data are presented as mean ± SD. One-way ANOVA was used for comparisons among multiple groups, *, P < 0.05; ns, not significant

Article Snippet: Following centrifugation, the supernatant was subjected to an overnight incubation with primary antibodies [rabbit anti-TAF7 (Cat# 13506-1-AP, Proteintech, USA) or rabbit anti-IgG (Cat# 10284-1-AP, Proteintech, USA)] at 4 °C with gentle rotation.

Techniques: Binding Assay, Negative Control, Luciferase, Quantitative RT-PCR, Expressing, Western Blot, Tube Formation Assay, Incubation, Flow Cytometry, Transwell Assay, Migration, Cell Counting, Wound Healing Assay

Journal: Biology Direct

Article Title: Single-cell RNA sequencing reveals that TAF7 promotes colorectal cancer stemness by targeting SOX2

doi: 10.1186/s13062-026-00910-4

Figure Lengend Snippet: In vivo validation of TAF7 knockdown on CRC cell stemness and EMT progression in mice. Mouse groups: sh-NC, sh-TAF7; A : Tumor volume in BALB/c Nude mice bearing xenografts after tumor cell injection, each group contained six independent animals ( n = 6); B-C : Weight ( B ) and representative images ( C ) of excised tumors, each group contained six independent animals ( n = 6); D : IHC staining for E-cadherin, N-cadherin, TAF7, SOX2, and CD133 in xenograft tumors. Scale bar = 50 μm, magnification: ×200, n = 3 independent experiments. In vivo data are presented as mean ± SD. Student’s t -test was used for comparisons between two groups, *, P < 0.05

Article Snippet: Following centrifugation, the supernatant was subjected to an overnight incubation with primary antibodies [rabbit anti-TAF7 (Cat# 13506-1-AP, Proteintech, USA) or rabbit anti-IgG (Cat# 10284-1-AP, Proteintech, USA)] at 4 °C with gentle rotation.

Techniques: In Vivo, Biomarker Discovery, Knockdown, Injection, Immunohistochemistry, Xenograft Assay