Journal: Scientific Reports

Article Title: A CXCR4 targeting peptide delivered by silica nanoparticles eliminates migrating cancer stem cells in pancreatic ductal adenocarcinoma

doi: 10.1038/s41598-026-48584-2

Figure Lengend Snippet: BMI1 downstream CXCL12–CXCR4 regulates EMT and stemness. ( A ) Protein–protein interactions of CXCL12 and CXCR4 with relevant factors involved in metastasis (red), stemness (purple), sonic hedgehog signaling (blue), AKT signaling (grey) and NFκB pathway (yellow) (STRING). ( B ) BMI1 gene expression analysis and western blot analysis. GAPDH was used as a loading control. Intensity ratios (IR) calculated against control lane using ImageJ. Cropped blot for clarity. ( C ) Gene expression analysis with genes involved in EMT using qRT-PCR. ( D ) Immunofluorescence quantifications and representative micrographs for indicated cell lines with white arrowheads marking mesenchymal structures of actin filaments stained with Phalloidin (pink) and nucleus stained with DAPI (blue). ( E ) Migration assays towards serum containing media. ( F ) Experimental scheme to evaluate CD133 and CXCR4 surface expression using flow cytometry for Panc354 and MetPO1 (sh_ SCR , sh1_ BMI1 and sh2_ BMI1 ) with (or without) CXCL12. ( G ) Flow cytometry analysis of CD133 + cells and CD133 + CXCR4 + cells in MetPO1 (sh_ SCR , sh1_ BMI1 and sh2_ BMI1 ) treated with (or without) CXCL12. ( H ) Representative cytometry plots for MetPO1 cell line. Error bars represent the standard deviation. n = 3 for all experiments unless otherwise depicted in the datasets. * p < 0.05, ns = not significant.

Article Snippet: Recombinant BMI1 IgG1 (REA438) , APC , Miltenyi Biotec , 130-124-301.

Techniques: Protein-Protein interactions, Gene Expression, Western Blot, Control, Quantitative RT-PCR, Immunofluorescence, Staining, Migration, Expressing, Flow Cytometry, Cytometry, Standard Deviation

Journal: Scientific Reports

Article Title: A CXCR4 targeting peptide delivered by silica nanoparticles eliminates migrating cancer stem cells in pancreatic ductal adenocarcinoma

doi: 10.1038/s41598-026-48584-2

Figure Lengend Snippet: JM#21, most potent EPIX4 derivative to target miCSCs. ( A ) Migration assays towards CXCL12 using Panc354 for EPI-X4 and its derivatives at depicted concentrations. Pre-treatment with EPI-X4, WSCO2, JM#21 and the inactive peptide was applied for 30 min. ( B ) Representative micrographs (10x, DAPI staining) of transwell migration assays in Panc354 cells for the indicated conditions and concentrations. ( C ) Migration assays towards CXCL12 for MetPO1 using JM#21 and the inactive peptide at depicted concentrations. ( D ) Quantifications of percent mesenchymal structures after 15 min and 6 h of CXCL12 treatment in MetPO1 cells. JM#21 pre-treatment was applied for 30 min and representative micrographs with white arrowheads marking mesenchymal structures of actin filaments stained with Phalloidin (pink) and nuclear staining using DAPI (blue). ( E ) Gene expression analysis for indicated cell lines with genes involved in EMT and SHH pathway. ( F ) Gene expression analysis for indicated cell lines with genes involved in stemness. ( G ) Sphere formation assays for 1 st and 2 nd generation of sphere formation. ( H ) Western blot analysis of CADHERIN-1, VIMENTIN, CADHERIN-2, NANOG and BMI1 for indicated cell lines. GAPDH was used as a loading control. Intensity ratios (IR) calculated against control lane using ImageJ. Cropped blot for clarity. ( I ) Experimental design for combination therapy analyzing relapse using JM#21, gemcitabine (labelled as G) and paclitaxel (labelled as P). Quantification of cell viability and representative pictures for clonogenic assays after treatment with JM#21 (10 μM ), gemcitabine (Gem) for indicated concentrations as depicted in experimental design in MetPO1 cell line. ( J ) Flow cytometry for CD133 in Panc354 and MetPO1cells for the indicated treatments shown as fold change. Error bars represent the standard deviation. n = 3 for all experiments unless otherwise depicted in the datasets. * p < 0.05, ns = not significant.

Article Snippet: Recombinant BMI1 IgG1 (REA438) , APC , Miltenyi Biotec , 130-124-301.

Techniques: Migration, Staining, Gene Expression, Western Blot, Control, Flow Cytometry, Standard Deviation

Journal: PLoS ONE

Article Title: Bmi1 Essentially Mediates Podocalyxin-Enhanced Cisplatin Chemoresistance in Oral Tongue Squamous Cell Carcinoma

doi: 10.1371/journal.pone.0123208

Figure Lengend Snippet: In (A) SCC-4 and (B) Tca8113 OTSCC cells, the protein levels of PODXL and BMI1 were determined with western blot analysis in normal control cells (NC, lane 1), cells stably transfected with the empty pcDNA 3.1 vector (VC, lane 2), cells stably transfected with PODXL (lane 3), cells stably transfected with PODXL and treated with focal adhesion kinase (FAK) inhibitor 14 (50 M) for 24 hours (PODXL+FAK-I, lane 4), cells stably transfected with Bmi1 (lane 5), cells stably transduced with scramble control shRNA (SC, lane 6), cells stably transduced with PODXL-shRNA (lane 7), cells stably transduced with BMI1-shRNA (lane 8), cells stably transfected with PODXL and transduced with BMI1-shRNA (PODXL+BMI1-shRNA, lane 9), and cells stably transfected with Bmi1 and transduced with PODXL-shRNA (Bmi1+PODXL-shRNA, lane 10). Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) blotting was used as a loading control. Density of the Western blots was measured by densitometry, and the density of the PODXL and the Bmi1 blots was normalized against that of the GAPHD blot in the same sample to obtain a relative blot density to represent relative PODXL and Bmi1 content in each sample, respectively. a p <0.05 vs. controls (NC, VC and SC); b p <0.05 vs. PODXL; c p <0.05 vs. PODXL+FAK-I; d p <0.05 vs. Bmi1; e p <0.05 vs. PODXL-shRNA; f p <0.05 vs. Bmi1-shRNA; g p <0.05 vs. PODXL+Bmi1-shRNA.

Article Snippet: Human Bmi1 cDNA clone (SC116894) was purchased from Origene (Beijing, China) and the full length Bmi1 cDNA sequence was subcloned into the pcDNA 3.1 plasmid.

Techniques: Western Blot, Control, Stable Transfection, Transfection, Plasmid Preparation, Transduction, shRNA

Journal: PLoS ONE

Article Title: Bmi1 Essentially Mediates Podocalyxin-Enhanced Cisplatin Chemoresistance in Oral Tongue Squamous Cell Carcinoma

doi: 10.1371/journal.pone.0123208

Figure Lengend Snippet: In (A) SCC-4 and (B) Tca8113 OTSCC cells, mRNA levels of Bmi1 were determined with real-time RT-PCR in normal control cells (NC), cells stably transfected with the empty pcDNA 3.1 vector (VC), cells stably transfected with PODXL, cells stably transfected with PODXL and treated with focal adhesion kinase (FAK) inhibitor 14 (50 M) for 24 hours (PODXL+FAK-I), cells stably transfected with Bmi1, cells stably transduced with scramble control shRNA (SC), cells stably transduced with PODXL-shRNA, cells stably transduced with BMI1-shRNA, cells stably transfected with PODXL and transduced with BMI1-shRNA (PODXL+BMI1-shRNA), and cells stably transfected with Bmi1 and transduced with PODXL-shRNA (Bmi1+PODXL-shRNA). a p <0.05 vs. controls (NC, VC and SC); b p <0.05 vs. PODXL; c p <0.05 vs. PODXL+FAK-I; d p <0.05 vs. Bmi1; e p <0.05 vs. PODXL-shRNA; f p <0.05 vs. Bmi1-shRNA; g p <0.05 vs. PODXL+Bmi1-shRNA.

Article Snippet: Human Bmi1 cDNA clone (SC116894) was purchased from Origene (Beijing, China) and the full length Bmi1 cDNA sequence was subcloned into the pcDNA 3.1 plasmid.

Techniques: Quantitative RT-PCR, Control, Stable Transfection, Transfection, Plasmid Preparation, Transduction, shRNA

Journal: PLoS ONE

Article Title: Bmi1 Essentially Mediates Podocalyxin-Enhanced Cisplatin Chemoresistance in Oral Tongue Squamous Cell Carcinoma

doi: 10.1371/journal.pone.0123208

Figure Lengend Snippet: (A) SCC-4 and (B) Tca8113 OTSCC cells were transfected with human Bmi1 promoter/luciferase reporter plasmids and then cultured for 24 hours. Luciferase activities were determined in normal control cells (NC), cells stably transfected with the empty pcDNA 3.1 vector (VC), cells stably transfected with PODXL, cells stably transfected with PODXL and treated with focal adhesion kinase (FAK) inhibitor 14 (50 M) for 24 hours (PODXL+FAK-I), cells stably transfected with Bmi1, cells stably transduced with scramble control shRNA (SC), cells stably transduced with PODXL-shRNA, cells stably transduced with BMI1-shRNA, cells stably transfected with PODXL and transduced with BMI1-shRNA (PODXL+BMI1-shRNA), and cells stably transfected with Bmi1 and transduced with PODXL-shRNA (Bmi1+PODXL-shRNA). The luciferase activity was expressed as fold changes to that of NC (designated as 1).

Article Snippet: Human Bmi1 cDNA clone (SC116894) was purchased from Origene (Beijing, China) and the full length Bmi1 cDNA sequence was subcloned into the pcDNA 3.1 plasmid.

Techniques: Transfection, Luciferase, Cell Culture, Control, Stable Transfection, Plasmid Preparation, Transduction, shRNA, Activity Assay

Journal: PLoS ONE

Article Title: Bmi1 Essentially Mediates Podocalyxin-Enhanced Cisplatin Chemoresistance in Oral Tongue Squamous Cell Carcinoma

doi: 10.1371/journal.pone.0123208

Figure Lengend Snippet: (A) SCC-4 and (B) Tca8113 OTSCC cells were pre-treated with transcription inhibitor actinomycin D (1 mg/mL) for 30 minutes, and then cultured for 1, 2 or 4 hours in medium containing actinomycin D (1 mg/mL). The Bmi1 mRNA level were then determined with real-time RT-PCR in normal control cells (NC), cells stably transfected with the empty pcDNA 3.1 vector (VC), cells stably transfected with PODXL, cells stably transfected with PODXL and treated with focal adhesion kinase (FAK) inhibitor 14 (50 M) for 24 hours (PODXL+FAK-I), cells stably transfected with Bmi1, cells stably transduced with scramble control shRNA (SC), cells stably transduced with PODXL-shRNA, cells stably transduced with BMI1-shRNA, cells stably transfected with PODXL and transduced with BMI1-shRNA (PODXL+BMI1-shRNA), and cells stably transfected with Bmi1 and transduced with PODXL-shRNA (Bmi1+PODXL-shRNA).

Article Snippet: Human Bmi1 cDNA clone (SC116894) was purchased from Origene (Beijing, China) and the full length Bmi1 cDNA sequence was subcloned into the pcDNA 3.1 plasmid.

Techniques: Cell Culture, Quantitative RT-PCR, Control, Stable Transfection, Transfection, Plasmid Preparation, Transduction, shRNA

Journal: PLoS ONE

Article Title: Bmi1 Essentially Mediates Podocalyxin-Enhanced Cisplatin Chemoresistance in Oral Tongue Squamous Cell Carcinoma

doi: 10.1371/journal.pone.0123208

Figure Lengend Snippet: (A) SCC-4 and (B) Tca8113 OTSCC cells were treated with or without various concentrations of cisplatin for 48 hours. The half maximal inhibitory concentration (IC50) values were determined in normal control cells (NC), cells stably transfected with the empty pcDNA 3.1 vector (VC), cells stably transfected with PODXL, cells stably transfected with PODXL and treated with focal adhesion kinase (FAK) inhibitor 14 (50 M) for 24 hours (PODXL+FAK-I), cells stably transfected with Bmi1, cells stably transduced with scramble control shRNA (SC), cells stably transduced with PODXL-shRNA, cells stably transduced with BMI1-shRNA, cells stably transfected with PODXL and transduced with BMI1-shRNA (PODXL+BMI1-shRNA), and cells stably transfected with Bmi1 and transduced with PODXL-shRNA (Bmi1+PODXL-shRNA). The IC50 dose-response curves were plotted with GraphPad Prism 5.0 (GraphPad Software). The dose-response curves for VC and SC in both SCC-4 and Tca8113 cells are presented in , because they mostly overlap with the dose-response curve of NC. IC50 values (mean±SD) are presented by histograms. a p <0.05 vs. controls (NC, VC and SC); b p <0.05 vs. PODXL; c p <0.05 vs. PODXL+FAK-I; d p <0.05 vs. Bmi1; e p <0.05 vs. PODXL-shRNA; f p <0.05 vs. Bmi1-shRNA; g p <0.05 vs. PODXL+Bmi1-shRNA.

Article Snippet: Human Bmi1 cDNA clone (SC116894) was purchased from Origene (Beijing, China) and the full length Bmi1 cDNA sequence was subcloned into the pcDNA 3.1 plasmid.

Techniques: Concentration Assay, Control, Stable Transfection, Transfection, Plasmid Preparation, Transduction, shRNA, Software

Journal: PLoS ONE

Article Title: Bmi1 Essentially Mediates Podocalyxin-Enhanced Cisplatin Chemoresistance in Oral Tongue Squamous Cell Carcinoma

doi: 10.1371/journal.pone.0123208

Figure Lengend Snippet: (A) SCC-4 and (B) Tca8113 OTSCC cells were treated with cisplatin (10 μM) for 12 and 24 hours. Apoptosis was measured with a microplate reader-based TiterTACS in situ apoptosis detection kit (R&D systems) in normal control cells (NC), cells stably transfected with the empty pcDNA 3.1 vector (VC), cells stably transfected with PODXL, cells stably transfected with PODXL and treated with focal adhesion kinase (FAK) inhibitor 14 (50 M) for 24 hours (PODXL+FAK-I), cells stably transfected with Bmi1, cells stably transduced with scramble control shRNA (SC), cells stably transduced with PODXL-shRNA, cells stably transduced with BMI1-shRNA, cells stably transfected with PODXL and transduced with BMI1-shRNA (PODXL+BMI1-shRNA), and cells stably transfected with Bmi1 and transduced with PODXL-shRNA (Bmi1+PODXL-shRNA). Cell apoptosis was shown as fold changes to that of untreated NC (at 0 hour of treatment; designated as 1). * p <0.05 vs. controls (NC, VC and SC).

Article Snippet: Human Bmi1 cDNA clone (SC116894) was purchased from Origene (Beijing, China) and the full length Bmi1 cDNA sequence was subcloned into the pcDNA 3.1 plasmid.

Techniques: In Situ, Control, Stable Transfection, Transfection, Plasmid Preparation, Transduction, shRNA

Journal: PLoS ONE

Article Title: Bmi1 Essentially Mediates Podocalyxin-Enhanced Cisplatin Chemoresistance in Oral Tongue Squamous Cell Carcinoma

doi: 10.1371/journal.pone.0123208

Figure Lengend Snippet: In (A) SCC-4 and (B) Tca8113 OTSCC cells, the FAK activity was determined with a Universal Tyrosine Kinase Assay kit (Takara Biomedical Technology) in normal control cells (NC), cells stably transfected with the empty pcDNA 3.1 vector (VC), cells stably transfected with PODXL, cells stably transfected with PODXL and treated with focal adhesion kinase (FAK) inhibitor 14 (50 M) for 24 hours (PODXL+FAK-I), cells stably transfected with Bmi1, cells stably transduced with scramble control shRNA (SC), cells stably transduced with PODXL-shRNA, cells stably transduced with BMI1-shRNA, cells stably transfected with PODXL and transduced with BMI1-shRNA (PODXL+BMI1-shRNA), and cells stably transfected with Bmi1 and transduced with PODXL-shRNA (Bmi1+PODXL-shRNA). The FAK activity was shown as fold changes to that of NC (designated as 1). a p <0.05 vs. controls (NC, VC and SC); b p <0.05 vs. PODXL; c p <0.05 vs. PODXL+FAK-I; d p <0.05 vs. Bmi1; e p <0.05 vs. PODXL-shRNA; f p <0.05 vs. Bmi1-shRNA; g p <0.05 vs. PODXL+Bmi1-shRNA.

Article Snippet: Human Bmi1 cDNA clone (SC116894) was purchased from Origene (Beijing, China) and the full length Bmi1 cDNA sequence was subcloned into the pcDNA 3.1 plasmid.

Techniques: Activity Assay, Universal Tyrosine Kinase Assay, Control, Stable Transfection, Transfection, Plasmid Preparation, Transduction, shRNA

Journal: Bone Research

Article Title: KDM6B safeguards mineralized tissue homeostasis from mechanical stress through epigenetic control of PIEZO1-mediated mechanotransduction in the mouse incisor

doi: 10.1038/s41413-026-00544-2

Figure Lengend Snippet: KDM6B epigenetically regulates BMI1 to silence PIEZO1 expression and maintain tissue homeostasis. a CUT&RUN profiles of H3K27me3 in the proximal incisor region of control and Gli1-CreER T2 ;Kdm6b fl/fl mice at 5 dpt. TSS: Transcription start sites. TES: Transcription end sites. b Venn diagram showing overlap between genes with increased H3K27me3 enrichment in CUT&RUN and downregulated genes in RNA-seq from Gli1-CreER T2 ;Kdm6b fl/fl incisors. c Heatmap of overlapping genes expressed in TACs in control and Gli1-CreER T2 ;Kdm6b fl/fl mice. d Co-staining of Bmi1 (white) and Kdm6b (red) in the wild-type mouse incisors. d’ Magnified view of boxed region. White dotted lines outline the cervical loop. Yellow arrows indicate double-positive cells. Scale bars, 50 μm. e CUT&RUN tracks showing H3K27me3 enrichment at the Bmi1 locus in control and Gli1-CreER T2 ;Kdm6b fl/fl incisors. IgG serves as a negative control. Red lines indicate CRISPRi gRNA target sites. f RT-qPCR analysis of Bmi1 expression following CRISPRi targeting (mean ± SEM, n = 3; gRNA1, P = 0.034 6; gRNA2, P = 0.709 1; gRNA3, P = 0.058 7). g –i Bmi1 in situ hybridization in control, Kdm6b mutant, and Ezh2 rescue incisors. g’ –i’ Magnified views of boxed regions. White dotted lines outline the cervical loop. Yellow arrows indicate positive signals. Asterisk indicates absence of signal. j Relative Bmi1 mRNA expression in the three genotypes (mean ± SEM, n = 3; control vs Kdm6b mutant, P = 0.004 2; Kdm6b mutant vs Ezh2 rescue, P = 0.000 2; control vs Ezh2 rescue, P = 0.012 8). k Piezo1 expression in dental mesenchymal cells after 3 days of control or Bmi1 siRNA treatment (mean ± SEM, n = 3; P = 0.036 4). l Piezo1 expression after vector or Bmi1 plasmid treatment (mean ± SEM, n = 3; ctrl+vector vs. ctrl+ Bmi1 , P = 0.022 6; ctrl+vector vs. mutant+vector, P = 0.000 8; ctrl+vector vs. mutant+ Bmi1 , P = 0.838 4; mutant+vector vs. mutant+ Bmi1 , P = 0.000 9). m CUT&RUN profiles showing BMI1 enrichment at the Piezo1 locus (IgG negative control and two BMI1 replicates). Red lines indicate CRISPRi gRNA target sites. n RT-qPCR analysis of Piezo1 expression following CRISPRi targeting of BMI1-binding regions (mean ± SEM, n = 3; gRNA1, P = 0.012 3 ; gRNA2, P = 0.029 7). Kdm6b mutant: Gli1-CreER T2 ;Kdm6b fl/fl . Ezh2 rescue: Gli1-CreER T2 ;Kdm6b fl/fl ;Ezh2 fl/+

Article Snippet: For plasmid transfection, cells were transfected with 1 μg/μL Bmi1 (Myc-DDK-tagged) plasmid (OriGene, MR226936; vector, PS100001 ) DNA using LipofectamineTM 3000 Transfection Reagent (Thermo Fisher Scientific, L3000150) for 3 days, followed by TUNEL staining and qPCR.

Techniques: Expressing, Control, RNA Sequencing, Staining, Negative Control, Quantitative RT-PCR, In Situ Hybridization, Mutagenesis, Plasmid Preparation, Binding Assay

Journal: Bone Research

Article Title: KDM6B safeguards mineralized tissue homeostasis from mechanical stress through epigenetic control of PIEZO1-mediated mechanotransduction in the mouse incisor

doi: 10.1038/s41413-026-00544-2

Figure Lengend Snippet: Schematic representation of KDM6B safeguarding tissue homeostasis to mechanical stress through epigenetic control of PIEZO1-mediated mechanotransduction. Using the mouse incisor as a model of mechanical loading, we reveal that within TACs, Kdm6b demethylates H3K27me3, thereby relieving the repression of the Bmi1 gene. Normal BMI1 inhibits Piezo1 expression. This maintains physiological PIEZO1 levels, ensuring calibrated Ca 2+ influx for proliferation and differentiation. In contrast, loss of Kdm6b leads to an accumulation of H3K27me3 at the Bmi1 promoter region, which silences Bmi1 expression and diminishes BMI1 formation. This reduction results in pathologically increased PIEZO1 ion channels in the membrane. The subsequent Ca 2+ overload triggers TAC apoptosis while reducing proliferation and differentiation. Ultimately, these molecular events compromise tissue homeostasis. Schematic created with BioRender.com. Ho, T. (2026) https://BioRender.com/8mzv4a3

Article Snippet: For plasmid transfection, cells were transfected with 1 μg/μL Bmi1 (Myc-DDK-tagged) plasmid (OriGene, MR226936; vector, PS100001 ) DNA using LipofectamineTM 3000 Transfection Reagent (Thermo Fisher Scientific, L3000150) for 3 days, followed by TUNEL staining and qPCR.

Techniques: Control, Expressing, Membrane

Journal: Cells

Article Title: CD80-Mediated T-Cell Suppression by Cancer Stem-like Cells in Head and Neck Squamous Cell Carcinoma

doi: 10.3390/cells15030266

Figure Lengend Snippet: CD80 is related to cancer-cell stemness in PR and CR patients. ( A ) UMAP plots show re-clustering of epithelial cells in PR and CR groups. ( B ) Pseudotime analysis demonstrates the derivation of epithelial cells in PR and CR groups based on the derived trajectory, tissue origin, and monocle states. ( C ) The Heatmap shows the top genes expressed with the pseudotime trajectory of three fate cells. ( D ) The Boxplot shows the stemness scores of three state cells. ( E ) The PR and CR proportions in each state. ( F ) The Dot plot shows the expression levels of stemness-related genes in each cluster of epithelial. ( G ) The Boxplot shows stemness scores among different clusters of epithelial cells. ( H ) The state proportion in each cluster. ( I ) Pseudotime analysis shows the derivation of clusters 0, 1, 4, 7, 9, and 10 from epithelial cells based on monocle states and cluster origin. ( J ) Trajectory of the expression of CD80. ( K ) Analysis of the correlation between CD80 and stemness-related genes ALDH1A1 , SOX9 , MET, and MYC in epithelial cells from PR and CR groups. ( L ) Analysis of the correlation between CD80 and stemness-related genes CD44 , BMI1 , MET, and POU5F1 in HNSCC based on TCGA HNSCC datasets.

Article Snippet: The primary antibodies used were anti-BMI1 antibody (Proteintech, Wuhan, China, Cat#66161-1-Ig) and anti-Caspase3 antibody (Absin Bioscience Inc., Shanghai, China, Cat#abs119676).

Techniques: Derivative Assay, Expressing

Journal: Cells

Article Title: CD80-Mediated T-Cell Suppression by Cancer Stem-like Cells in Head and Neck Squamous Cell Carcinoma

doi: 10.3390/cells15030266

Figure Lengend Snippet: CD80 is highly expressed in CSCs. ( A ) Western blot shows protein expression levels of CD80 in 10 paired HNSCC tissues. ( B ) The expression level of CD80 in normal epithelial cell line (HaCaT) and HNSCC cell lines (HN6, CAL27, SCC9, and SCC25). Values are mean ± SD. ns, no significance; ** p < 0.01; *** p < 0.001; One-way ANOVA test; n = 3. ( C ) Flow cytometry isolates ALDH high CD44 + populations from HNSCC cell lines. The left panel shows the fluorescence minus one (FMO) control (the ALDH-only control). And RT-qPCR shows significantly higher CD80 expression in ALDH high CD44 + versus ALDH low CD44 − cancer cells. Values are mean ± SD. * p < 0.05; *** p < 0.001; Student’s t test, n = 3. ( D ) Flow cytometry isolates BMI1 + populations from HNSCC cell lines, and RT-qPCR shows significantly higher CD80 expression in BMI1 + versus BMI1 − cancer cells. Values are mean ± SD. * p < 0.05; *** p < 0.001; Student’s t test, n = 3. ( E ) RT-qPCR shows significantly higher CD80 expression in tumor sphere versus adherent cells in HNSCC cells. Values are mean ± SD. *** p < 0.001; Student’s t test, n = 3. ( F ) Representative images of flow-cytometry show CD80 + HNSCC cells. ( G ) Representative images of tumor spheres and quantification of the sphere number of CD80 − and CD80 + HNSCC cells. Scale bar, 200 μm. Values are mean ± SD. * p < 0.05; ** p < 0.01; Student’s t test, n = 3.

Article Snippet: The primary antibodies used were anti-BMI1 antibody (Proteintech, Wuhan, China, Cat#66161-1-Ig) and anti-Caspase3 antibody (Absin Bioscience Inc., Shanghai, China, Cat#abs119676).

Techniques: Western Blot, Expressing, Flow Cytometry, Fluorescence, Control, Quantitative RT-PCR