Journal: Journal of Hepatocellular Carcinoma

Article Title: CENPF Overexpression Induced by HBV Infection Facilitates the G1/S Cell Cycle Transition of Hepatocellular Carcinoma Cells via MYC Pathway

doi: 10.2147/JHC.S580622

Figure Lengend Snippet: The expression of CENPF was higher in HCC tissues with HBV infection. ( A ) Representative immunohistochemical images of CENPF expression in HCC and adjacent tissues (n=100). ( B ) CENPF mRNA expression in HCC and adjacent tissues was examined by real-time PCR (n=27 pairs). ( C ) Immunohistochemical analysis showed that the positive rate of CENPF in poorly differentiated HCC tissues was higher than that in well-differentiated or moderately differentiated HCC tissues (n=100). ( D and E ) CENPF expression in HBV-negative and -positive HCC tissues was measured by qRT-PCR (n=27 pairs) and immunohistochemical analysis (n=100). (*p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001).

Article Snippet: Briefly, each reaction mixture contained 2X TaqMan TM Genotyping Master Mix (4371353, Applied Biosystems, USA), 20X TaqMan TM Copy Number Reference Assay reagent (RNase P, 4403326, Applied Biosystems, USA), 20X TaqMan TM Copy Number Assay reagent ( CENPF , Hs03027722_cn, Applied Biosystems, USA) and template DNA.

Techniques: Expressing, Infection, Immunohistochemical staining, Real-time Polymerase Chain Reaction, Quantitative RT-PCR

Journal: Journal of Hepatocellular Carcinoma

Article Title: CENPF Overexpression Induced by HBV Infection Facilitates the G1/S Cell Cycle Transition of Hepatocellular Carcinoma Cells via MYC Pathway

doi: 10.2147/JHC.S580622

Figure Lengend Snippet: The frequency of CENPF amplification was higher in HBV-related HCC and HBx enhanced the expression of CENPF in HCC. ( A and B ) The amplification frequency of CENPF in HBV-positive HCC tissues (n=39) was higher than that in HBV-negative HCC tissues (n=19). ( C ) CENPF copy number was positively correlated with CENPF mRNA expression levels (n=39). ( D and E ) The effects of transient HBx overexpression in HCCLM3 and Hep3B cell lines on the mRNA and protein expression levels of CENPF. Data are mean ± SD of three independent experiments. (**p < 0.01, ****p < 0.0001).

Article Snippet: Briefly, each reaction mixture contained 2X TaqMan TM Genotyping Master Mix (4371353, Applied Biosystems, USA), 20X TaqMan TM Copy Number Reference Assay reagent (RNase P, 4403326, Applied Biosystems, USA), 20X TaqMan TM Copy Number Assay reagent ( CENPF , Hs03027722_cn, Applied Biosystems, USA) and template DNA.

Techniques: Amplification, Expressing, Over Expression

Journal: Journal of Hepatocellular Carcinoma

Article Title: CENPF Overexpression Induced by HBV Infection Facilitates the G1/S Cell Cycle Transition of Hepatocellular Carcinoma Cells via MYC Pathway

doi: 10.2147/JHC.S580622

Figure Lengend Snippet: Effects of CENPF knockdown and CENPF overexpression on HCC cell proliferation. ( A ) Western blotting analysis of CENPF expression in HCC cell lines. ( B ) CENPF knockdown was confirmed by Western blotting and qRT-PCR. Data were mean±SD of three independent experiments. ( C ) CENPF overexpression cell models were successfully constructed using the CRISPR/dCas9 SAM system. The SAM system relies on sgRNA to target specific sequences. Upon binding to the target sequence, the transcriptional activation system recruits the transcription initiation complex to activate the expression of the target gene. Data were mean±SD of three independent experiments. ( D and E ) The proliferation rates of HCC cell lines with CENPF knockdown or CENPF overexpression were analyzed by CCK-8. Data were mean±SD of four replicate wells. ( F and H ) Photos and weight analyses of xenografts excised from nude mice inoculated with CENPF-knockdown (n = 3/group) or CENPF-overexpressing (n = 6/group) HCC cell lines. ( G and I ) Representative images of CENPF IHC staining in nude mice tumor tissues. (*p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001).

Article Snippet: Briefly, each reaction mixture contained 2X TaqMan TM Genotyping Master Mix (4371353, Applied Biosystems, USA), 20X TaqMan TM Copy Number Reference Assay reagent (RNase P, 4403326, Applied Biosystems, USA), 20X TaqMan TM Copy Number Assay reagent ( CENPF , Hs03027722_cn, Applied Biosystems, USA) and template DNA.

Techniques: Knockdown, Over Expression, Western Blot, Expressing, Quantitative RT-PCR, Construct, CRISPR, Binding Assay, Sequencing, Activation Assay, CCK-8 Assay, Immunohistochemistry

Journal: Journal of Hepatocellular Carcinoma

Article Title: CENPF Overexpression Induced by HBV Infection Facilitates the G1/S Cell Cycle Transition of Hepatocellular Carcinoma Cells via MYC Pathway

doi: 10.2147/JHC.S580622

Figure Lengend Snippet: Effects of CENPF knockdown and CENPF overexpression on the HCC cell cycle. ( A and B ) The cell cycle distributions of CENPF knockdown Huh-7 and Hep3B cell lines with relatively high/medium endogenous CENPF expression. ( C and D ) The cell cycle distributions of CENPF overexpression Huh-7 and Hep3B cell lines with relatively low/medium endogenous CENPF expression. The cell cycle distributions were determined by flow cytometry. Data were mean±SD of three independent experiments. (**p < 0.01, ***p < 0.001).

Article Snippet: Briefly, each reaction mixture contained 2X TaqMan TM Genotyping Master Mix (4371353, Applied Biosystems, USA), 20X TaqMan TM Copy Number Reference Assay reagent (RNase P, 4403326, Applied Biosystems, USA), 20X TaqMan TM Copy Number Assay reagent ( CENPF , Hs03027722_cn, Applied Biosystems, USA) and template DNA.

Techniques: Knockdown, Over Expression, Expressing, Flow Cytometry

Journal: Journal of Hepatocellular Carcinoma

Article Title: CENPF Overexpression Induced by HBV Infection Facilitates the G1/S Cell Cycle Transition of Hepatocellular Carcinoma Cells via MYC Pathway

doi: 10.2147/JHC.S580622

Figure Lengend Snippet: Transcriptome analysis of HCC cells with CENPF knockdown and CENPF overexpression respectively. ( A and C ) Significantly enriched hallmark gene sets analyzed by GSEA based on the RNA-seq data. ( B and D ) GSEA plots showed significantly enriched proliferation-related hallmarks. ( E and F ) Validation of the RNA-seq analysis by qRT-PCR. The red colored dotted rectangular boxes highlighted the expressions of the MYC gene as detected by real-time PCR in the CENPF knockdown Huh-7 cell line and the CENPF overexpression HCCLM3 cell line. ( G ) Flow cytometry was used to analyze the effect of the MYC pathway inhibitor 10058-F4 on the cell cycle of CENPF-overexpressing HCCLM3 cells. Data were mean±SD of three independent experiments. (*p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001).

Article Snippet: Briefly, each reaction mixture contained 2X TaqMan TM Genotyping Master Mix (4371353, Applied Biosystems, USA), 20X TaqMan TM Copy Number Reference Assay reagent (RNase P, 4403326, Applied Biosystems, USA), 20X TaqMan TM Copy Number Assay reagent ( CENPF , Hs03027722_cn, Applied Biosystems, USA) and template DNA.

Techniques: Knockdown, Over Expression, RNA Sequencing, Biomarker Discovery, Quantitative RT-PCR, Real-time Polymerase Chain Reaction, Flow Cytometry

Journal: Journal of Hepatocellular Carcinoma

Article Title: CENPF Overexpression Induced by HBV Infection Facilitates the G1/S Cell Cycle Transition of Hepatocellular Carcinoma Cells via MYC Pathway

doi: 10.2147/JHC.S580622

Figure Lengend Snippet: A schematic model illustrating the procedure of HBV induced CENPF overexpression in the proliferation of HCC cells. HBV integrates into human genome, lead to genomic instability and the CENPF gene amplification, resulted in the upregulated CENPF expression in HCC. The CENPF overexpression promotes HCC proliferation by facilitating G1/S transition via the MYC pathway.

Article Snippet: Briefly, each reaction mixture contained 2X TaqMan TM Genotyping Master Mix (4371353, Applied Biosystems, USA), 20X TaqMan TM Copy Number Reference Assay reagent (RNase P, 4403326, Applied Biosystems, USA), 20X TaqMan TM Copy Number Assay reagent ( CENPF , Hs03027722_cn, Applied Biosystems, USA) and template DNA.

Techniques: Over Expression, Amplification, Expressing

Journal: Frontiers in Immunology

Article Title: Generation and characterization of human induced pluripotent stem cells from neuropathologically confirmed multiple system atrophy patient-derived fibroblasts

doi: 10.3389/fimmu.2026.1641981

Figure Lengend Snippet: Assessment of the cell line identity and the genomic integrity of the six MSA patient-derived iPSCs. (A) Short tandem repeat profiling provided the number of repeating units of 16 polymorphic microsatellite loci, to compare the donor fibroblast cells (reference) and the generated iPSCs and verify their origin. (B) G-banding revealed that the six iPSC lines showed a normal diploid karyotype (n = 46) and matched the sex of the donor patient. (C) Digital PCR allowed us to screen whether specific genomic regions accumulated duplications that may give a strong selective survival or growth advantage to iPSCs. Example of the scatter digital PCR plots displaying simultaneously (1) FAM signals, shown in purple and green to detect the target gene ( ID1 , NCAPD2 , PITX1 , RPS6KB1 , SOAT1 , or STS ), which are located in the 6 most recurrent abnormal regions; and (2) VIM signals, shown in orange and green to detect the reference gene, RPP30 . The ratio between the number of copies of the target gene and of the reference gene was used to determine the normal 2-copy number or copy number variations. The plots were obtained through QuantStudio 3D Digital PCR system. In all cases, we obtained 2-copy number (D) Quantitative Real Time RT-PCR was used for confirming the absence of the Sendai virus in the iPSC lines after 8 passages, through four primer sets (SeV, KOS, Klf4 and c-Myc). Box and Whisker plots depict in log10 scale the gene expression relative to GAPDH gene and normalized to the 7 days post-transduction fibroblasts, calculated by the 2(-ΔΔC(T)) method. In all iPSC clones, the expression levels of the SeV RNA were very low, reaching the limit of detection. Data are shown as the mean ± s.d. n = 3.

Article Snippet: STS , Hs00091141_cn , FAM , Xp22 , hg38|7350117-7350227 , 110 bp , MseI, HindIII, CviQI.

Techniques: Derivative Assay, Generated, Digital PCR, Quantitative RT-PCR, Virus, Whisker Assay, Gene Expression, Transduction, Clone Assay, Expressing