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The profiles and identification of differentially expressed circRNAs in cervical squamous-cell carcinoma and precursor tissues. A The box plot of the FPKM of 24 sequencing samples. B The numbers of differentially expressed circRNAs between two groups. C – H Volcano plots represented differentially expressed circRNAs between two groups. The red and blue dots represent up- and down- regulated circRNAs, respectively, by at least onefold significantly (P-values < 0.05). I Overlap of upregulated circRNAs in HSIL and CSCC group compared with HPV-NCE group. J Overlap of upregulated circRNAs in HSIL and CSCC group compared with HPV16+ NCE group. K – M Relative expression levels of circRP13-279N23.2, circSTAG1 and <t>circPOLD1</t> measured by qRT-PCR in HPV-NCE (n = 32), HPV16+ HSIL (n = 19) and HPV16+ CSCC (n = 22) group. In each assay, the scatter plot represents median with interquartile range (*P < 0.05, **P < 0.01, ***P < 0.001, Kruskal–Wallis test). N – P Relative expression levels of circRP13-279N23.2, circSTAG1, and circPOLD1 measured by qRT-PCR in HPV-NCE (n = 32), HSIL with all-type HPV positive (n = 46), and CSCC with all-type HPV positive (n = 48) group. In each assay, the scatter plot represents median with interquartile range (*P < 0.05, **P < 0.01, ***P < 0.001, Kruskal–Wallis test)
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Bioinformatics analysis and screening of differentially expressed genes between wild-type control and BL-I PF lung tissues of mice in the gene expression omnibus database. (a) The heat map and volcano plot of differentiating genes in the GSE43695 dataset. (b) The heat map and volcano plot of differentiating genes in the GSE123293 dataset. (c) The Venn diagram illustrating the intersection of low-expression differential genes in the GSE123293 and GSE43695 datasets. (d) The Venn diagram illustrating the intersection of high-expression differential genes in the GSE123293 and GSE43695 datasets. (e) Intersection of highly expressed distinguishing genes in the GSE123293 and GSE43695 datasets (difference multiple >2). <t>Zbtb16:</t> Zinc finger and BTB domain containing 16, HPGD: 15-hydroxyprostaglandin dehydrogenase, AOX3: Aldehyde oxidase 3, DBP: D-box binding PAR BZIP transcription factor, S100A8: S100 calcium binding protein A8, S100A9: S100 calcium binding protein A9, BL-I: Bleomycin-induced, PF: Pulmonary fibrosis.
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The profiles and identification of differentially expressed circRNAs in cervical squamous-cell carcinoma and precursor tissues. A The box plot of the FPKM of 24 sequencing samples. B The numbers of differentially expressed circRNAs between two groups. C – H Volcano plots represented differentially expressed circRNAs between two groups. The red and blue dots represent up- and down- regulated circRNAs, respectively, by at least onefold significantly (P-values < 0.05). I Overlap of upregulated circRNAs in HSIL and CSCC group compared with HPV-NCE group. J Overlap of upregulated circRNAs in HSIL and CSCC group compared with HPV16+ NCE group. K – M Relative expression levels of circRP13-279N23.2, circSTAG1 and circPOLD1 measured by qRT-PCR in HPV-NCE (n = 32), HPV16+ HSIL (n = 19) and HPV16+ CSCC (n = 22) group. In each assay, the scatter plot represents median with interquartile range (*P < 0.05, **P < 0.01, ***P < 0.001, Kruskal–Wallis test). N – P Relative expression levels of circRP13-279N23.2, circSTAG1, and circPOLD1 measured by qRT-PCR in HPV-NCE (n = 32), HSIL with all-type HPV positive (n = 46), and CSCC with all-type HPV positive (n = 48) group. In each assay, the scatter plot represents median with interquartile range (*P < 0.05, **P < 0.01, ***P < 0.001, Kruskal–Wallis test)

Journal: Journal of Translational Medicine

Article Title: The biomarker potential of circPOLD1 and its binding protein YBX1 in cervical carcinogenesis

doi: 10.1186/s12967-025-06494-3

Figure Lengend Snippet: The profiles and identification of differentially expressed circRNAs in cervical squamous-cell carcinoma and precursor tissues. A The box plot of the FPKM of 24 sequencing samples. B The numbers of differentially expressed circRNAs between two groups. C – H Volcano plots represented differentially expressed circRNAs between two groups. The red and blue dots represent up- and down- regulated circRNAs, respectively, by at least onefold significantly (P-values < 0.05). I Overlap of upregulated circRNAs in HSIL and CSCC group compared with HPV-NCE group. J Overlap of upregulated circRNAs in HSIL and CSCC group compared with HPV16+ NCE group. K – M Relative expression levels of circRP13-279N23.2, circSTAG1 and circPOLD1 measured by qRT-PCR in HPV-NCE (n = 32), HPV16+ HSIL (n = 19) and HPV16+ CSCC (n = 22) group. In each assay, the scatter plot represents median with interquartile range (*P < 0.05, **P < 0.01, ***P < 0.001, Kruskal–Wallis test). N – P Relative expression levels of circRP13-279N23.2, circSTAG1, and circPOLD1 measured by qRT-PCR in HPV-NCE (n = 32), HSIL with all-type HPV positive (n = 46), and CSCC with all-type HPV positive (n = 48) group. In each assay, the scatter plot represents median with interquartile range (*P < 0.05, **P < 0.01, ***P < 0.001, Kruskal–Wallis test)

Article Snippet: Small interference RNAs (siRNAs) that target the back-spliced junction of circPOLD1 were designed and synthesized by GenePharma (Shanghai, China).

Techniques: Sequencing, Expressing, Quantitative RT-PCR

Identification of circPOLD1 as a circular RNA. A The genomic loci of POLD1 gene and circPOLD1. Sanger sequencing of qRT-PCR product of circPOLD1 showed the back-spliced junction. B The qRT-PCR analysis of circPOLD1 after treated with RNase R in SiHa and CaSki cells (***P < 0.001, Student’s t-test). C CircPOLD1 was validated by RT-PCR using divergent and convergent primers in complementary DNA (cDNA) and genomic DNA (gDNA) in SiHa cells. D The sequence of circPOLD1 was obtained from sanger sequencing (left). PCR products of circPOLD1 with two pairs of divergent primers that amplified the whole sequence of circPOLD1 in SiHa cells (right). E The circPOLD1 expression measured by Northern blot in RNA of SiHa cells treated with or without RNase R digestion, respectively. Probe of β-actin was incubated in the same membrane as control. F The circPOLD1 subcellular localization in SiHa and CaSki cells measured by fluorescent in situ hybridization assay. CircPOLD1 was labeled with a specific probe (red) for the back-splice junction, and the nucleus was labeled with DAPI (blue)

Journal: Journal of Translational Medicine

Article Title: The biomarker potential of circPOLD1 and its binding protein YBX1 in cervical carcinogenesis

doi: 10.1186/s12967-025-06494-3

Figure Lengend Snippet: Identification of circPOLD1 as a circular RNA. A The genomic loci of POLD1 gene and circPOLD1. Sanger sequencing of qRT-PCR product of circPOLD1 showed the back-spliced junction. B The qRT-PCR analysis of circPOLD1 after treated with RNase R in SiHa and CaSki cells (***P < 0.001, Student’s t-test). C CircPOLD1 was validated by RT-PCR using divergent and convergent primers in complementary DNA (cDNA) and genomic DNA (gDNA) in SiHa cells. D The sequence of circPOLD1 was obtained from sanger sequencing (left). PCR products of circPOLD1 with two pairs of divergent primers that amplified the whole sequence of circPOLD1 in SiHa cells (right). E The circPOLD1 expression measured by Northern blot in RNA of SiHa cells treated with or without RNase R digestion, respectively. Probe of β-actin was incubated in the same membrane as control. F The circPOLD1 subcellular localization in SiHa and CaSki cells measured by fluorescent in situ hybridization assay. CircPOLD1 was labeled with a specific probe (red) for the back-splice junction, and the nucleus was labeled with DAPI (blue)

Article Snippet: Small interference RNAs (siRNAs) that target the back-spliced junction of circPOLD1 were designed and synthesized by GenePharma (Shanghai, China).

Techniques: Sequencing, Quantitative RT-PCR, Reverse Transcription Polymerase Chain Reaction, Amplification, Expressing, Northern Blot, Incubation, Membrane, Control, In Situ Hybridization, Labeling

CircPOLD1 promotes the proliferation through directly binding to YBX1 in cervical cancer cells. A Schematic representation of the sites of the siRNAs specific to the back-splice junction of circPOLD1. B The circPOLD1 and POLD1 mRNA expression measured by qRT-PCR analysis in SiHa and CaSki cells after transfected with two siRNAs. Data were the means ± SEM of three independent experiments (*P < 0.05, **P < 0.01, ***P < 0.001, Student’s t-test). C The proliferation measured by CCK-8 assay in SiHa and CaSki cells transfected with circPOLD1 siRNAs, oligo control (oligo) and without treated (wildtype). Data were the means ± SEM of three experiments (*P < 0.05, **P < 0.01, ***P < 0.001, Student’s t-test). D Representative image (left) and quantification (right panel) of colony formation assays in SiHa and CaSki cells transfected with circPOLD1 siRNAs or oligo control. Data were the means ± SEM of three experiments (*P < 0.05, **P < 0.01, Student’s t-test). E Apoptotic rates measured by flow cytometry in SiHa and CaSki cells transfected with circPOLD1 siRNAs or oligo control. Data were the means ± SEM of five experiments (*P < 0.05, **P < 0.01, Student’s t-test). F The circPOLD1 expression measured by qRT-PCR in SiHa cells after stably transfected with circPOLD1 shRNA(sh-circPOLD1) or oligo shRNA(oligo). G SiHa cells with stably transfected sh-circPOLD1 or oligo were subcutaneously injected into 6 BALB/c nude mice (4–6 weeks old, female), respectively, to establish the xenograft models. Top panel: The growths of xenografts were compared between sh-circPOLD1 and oligo group (*P < 0.05, **P < 0.01, ***P < 0.001, Student’s t-test). Tumor volumes were measured per week. Bottom panel: The photo showed the sizes of tumors when mice were sacrificed at week 8. H YBX1, the circPOLD1 binding protein predicted by online web server RBPmap. I , J Probes targeting back-splice junction of circPOLD1 and control oligo were transcribed in vitro, biotinylated, captured by streptavidin magnetic beads, and incubated with CaSki whole-cell lysates for RNA pull-down assays. The expression level of RNA measured by qRT-PCR in the eluate (left). CircPLD1 specific bands were assessed by Western blotting (right). Data were the means ± SEM of three experiments (*P < 0.05, Student’s t-test). K RIP assays were performed using antibodies against YBX1 in CaSki cells. The quantity of circPOLD1 was evaluated by qRT-PCR. Data were the means ± SEM of three experiments (*P < 0.05, Student’s t-test). L The expression level of YBX1 protein in SiHa cells after treated with two YBX1 siRNAs. M The proliferation by CCK-8 assay in SiHa and CaSki cells transfected with oligo control or YBX1 siRNAs. Data were the means ± SEM of three experiments (*P < 0.05, **P < 0.01, One-way ANOVA). N Representative images (left) and quantification (right) of colony formation assays in SiHa and CaSki cells transfected with YBX1 siRNAs or control. Data were the means ± SEM of three experiments (*P < 0.05, **P < 0.01, One-way ANOVA). O Co-transfection with circPOLD1 siRNA (si-circPOLD1) and YBX1 plasmid in CaSki cells was established. Down-regulated circPOLD1 expression and up-regulated YBX1 expression were evaluated by qRT-PCR. P Relative mRNA expressions of three downstream target genes of YBX1 (CDC25A, CIAPIN1 and XIAP) were measured by qRT-PCR in CaSki cells treated with control vector, si-circPOLD1, and si-circPOLD1 plus YBX1, respectively. Q The proliferation measured by CCK8 assay in CaSki cells treated with control vector, si-circPOLD1, and si-circPOLD1 plus YBX1, respectively. R Representative images of colony formation assays in CaSki cells treated with control vector, si-circPOLD1 and si-circPOLD1 plus YBX1, respectively. Data were the means ± SEM of three experiments (*P < 0.05, **P < 0.01, One-way ANOVA)

Journal: Journal of Translational Medicine

Article Title: The biomarker potential of circPOLD1 and its binding protein YBX1 in cervical carcinogenesis

doi: 10.1186/s12967-025-06494-3

Figure Lengend Snippet: CircPOLD1 promotes the proliferation through directly binding to YBX1 in cervical cancer cells. A Schematic representation of the sites of the siRNAs specific to the back-splice junction of circPOLD1. B The circPOLD1 and POLD1 mRNA expression measured by qRT-PCR analysis in SiHa and CaSki cells after transfected with two siRNAs. Data were the means ± SEM of three independent experiments (*P < 0.05, **P < 0.01, ***P < 0.001, Student’s t-test). C The proliferation measured by CCK-8 assay in SiHa and CaSki cells transfected with circPOLD1 siRNAs, oligo control (oligo) and without treated (wildtype). Data were the means ± SEM of three experiments (*P < 0.05, **P < 0.01, ***P < 0.001, Student’s t-test). D Representative image (left) and quantification (right panel) of colony formation assays in SiHa and CaSki cells transfected with circPOLD1 siRNAs or oligo control. Data were the means ± SEM of three experiments (*P < 0.05, **P < 0.01, Student’s t-test). E Apoptotic rates measured by flow cytometry in SiHa and CaSki cells transfected with circPOLD1 siRNAs or oligo control. Data were the means ± SEM of five experiments (*P < 0.05, **P < 0.01, Student’s t-test). F The circPOLD1 expression measured by qRT-PCR in SiHa cells after stably transfected with circPOLD1 shRNA(sh-circPOLD1) or oligo shRNA(oligo). G SiHa cells with stably transfected sh-circPOLD1 or oligo were subcutaneously injected into 6 BALB/c nude mice (4–6 weeks old, female), respectively, to establish the xenograft models. Top panel: The growths of xenografts were compared between sh-circPOLD1 and oligo group (*P < 0.05, **P < 0.01, ***P < 0.001, Student’s t-test). Tumor volumes were measured per week. Bottom panel: The photo showed the sizes of tumors when mice were sacrificed at week 8. H YBX1, the circPOLD1 binding protein predicted by online web server RBPmap. I , J Probes targeting back-splice junction of circPOLD1 and control oligo were transcribed in vitro, biotinylated, captured by streptavidin magnetic beads, and incubated with CaSki whole-cell lysates for RNA pull-down assays. The expression level of RNA measured by qRT-PCR in the eluate (left). CircPLD1 specific bands were assessed by Western blotting (right). Data were the means ± SEM of three experiments (*P < 0.05, Student’s t-test). K RIP assays were performed using antibodies against YBX1 in CaSki cells. The quantity of circPOLD1 was evaluated by qRT-PCR. Data were the means ± SEM of three experiments (*P < 0.05, Student’s t-test). L The expression level of YBX1 protein in SiHa cells after treated with two YBX1 siRNAs. M The proliferation by CCK-8 assay in SiHa and CaSki cells transfected with oligo control or YBX1 siRNAs. Data were the means ± SEM of three experiments (*P < 0.05, **P < 0.01, One-way ANOVA). N Representative images (left) and quantification (right) of colony formation assays in SiHa and CaSki cells transfected with YBX1 siRNAs or control. Data were the means ± SEM of three experiments (*P < 0.05, **P < 0.01, One-way ANOVA). O Co-transfection with circPOLD1 siRNA (si-circPOLD1) and YBX1 plasmid in CaSki cells was established. Down-regulated circPOLD1 expression and up-regulated YBX1 expression were evaluated by qRT-PCR. P Relative mRNA expressions of three downstream target genes of YBX1 (CDC25A, CIAPIN1 and XIAP) were measured by qRT-PCR in CaSki cells treated with control vector, si-circPOLD1, and si-circPOLD1 plus YBX1, respectively. Q The proliferation measured by CCK8 assay in CaSki cells treated with control vector, si-circPOLD1, and si-circPOLD1 plus YBX1, respectively. R Representative images of colony formation assays in CaSki cells treated with control vector, si-circPOLD1 and si-circPOLD1 plus YBX1, respectively. Data were the means ± SEM of three experiments (*P < 0.05, **P < 0.01, One-way ANOVA)

Article Snippet: Small interference RNAs (siRNAs) that target the back-spliced junction of circPOLD1 were designed and synthesized by GenePharma (Shanghai, China).

Techniques: Binding Assay, Expressing, Quantitative RT-PCR, Transfection, CCK-8 Assay, Control, Flow Cytometry, Stable Transfection, shRNA, Injection, In Vitro, Magnetic Beads, Incubation, Western Blot, Cotransfection, Plasmid Preparation

CircPOLD1 regulates AKT/mTOR/HIF-1α and glycolysis signaling pathway through phosphorylating YBX1 in cervical cancer cells. A , B Left panel: The YBX1 and p-YBX1 protein expression in CaSki and SiHa cells transfected with circPOLD1 siRNAs or control through western blot. Right panel: The quantitative analysis of YBX1 and p-YBX1 protein expression by ImageJ (**P < 0.01, ***P < 0.001, One-way ANOVA). C Venn diagram showing the overlap differentially genes between si-YBX1 vs si-NC and si-circPOLD1 vs si-NC via RNA sequencing. D The cluster heatmap of overlap differentially expressed mRNAs in YBX1 and circPOLD1 knockdown or control cells. E The pathway enrichment analysis of overlapping differential genes between si-YBX1 vs si-NC and si-circPOLD1 vs si-NC via RNA sequencing. F Protein levels of YBX1, p-YBX1, LDHA, HK2, HIF-1α, mTOR, p-mTOR, AKT and p-AKT, were determined following transfection with two YBX1 siRNAs and si-NC in CaSki and SiHa cells. G The quantitative analysis of F by ImageJ (*P < 0.05, **P < 0.01, ***P < 0.001, One-way ANOVA). H Protein levels of LDHA, HK2, HIF-1α, mTOR, p-mTOR, AKT and p-AKT were determined following transfection with two circPOLD1 siRNAs and si-NC in CaSki and SiHa cells. I The quantitative analysis of H by ImageJ (**P < 0.01, ***P < 0.001, One-way ANOVA)

Journal: Journal of Translational Medicine

Article Title: The biomarker potential of circPOLD1 and its binding protein YBX1 in cervical carcinogenesis

doi: 10.1186/s12967-025-06494-3

Figure Lengend Snippet: CircPOLD1 regulates AKT/mTOR/HIF-1α and glycolysis signaling pathway through phosphorylating YBX1 in cervical cancer cells. A , B Left panel: The YBX1 and p-YBX1 protein expression in CaSki and SiHa cells transfected with circPOLD1 siRNAs or control through western blot. Right panel: The quantitative analysis of YBX1 and p-YBX1 protein expression by ImageJ (**P < 0.01, ***P < 0.001, One-way ANOVA). C Venn diagram showing the overlap differentially genes between si-YBX1 vs si-NC and si-circPOLD1 vs si-NC via RNA sequencing. D The cluster heatmap of overlap differentially expressed mRNAs in YBX1 and circPOLD1 knockdown or control cells. E The pathway enrichment analysis of overlapping differential genes between si-YBX1 vs si-NC and si-circPOLD1 vs si-NC via RNA sequencing. F Protein levels of YBX1, p-YBX1, LDHA, HK2, HIF-1α, mTOR, p-mTOR, AKT and p-AKT, were determined following transfection with two YBX1 siRNAs and si-NC in CaSki and SiHa cells. G The quantitative analysis of F by ImageJ (*P < 0.05, **P < 0.01, ***P < 0.001, One-way ANOVA). H Protein levels of LDHA, HK2, HIF-1α, mTOR, p-mTOR, AKT and p-AKT were determined following transfection with two circPOLD1 siRNAs and si-NC in CaSki and SiHa cells. I The quantitative analysis of H by ImageJ (**P < 0.01, ***P < 0.001, One-way ANOVA)

Article Snippet: Small interference RNAs (siRNAs) that target the back-spliced junction of circPOLD1 were designed and synthesized by GenePharma (Shanghai, China).

Techniques: Expressing, Transfection, Control, Western Blot, RNA Sequencing, Knockdown

The biomarker potential of circPOLD1 in cervical carcinogenesis. A Reprehensive images of BaseScope staining of circPOLD1 (red arrows) and immunohistochemical staining of YBX1 expression in NCE, LSIL, HSIL, and CSCC tissues. The scale bar represents 50 um. B Statistics analysis of density of circPOLD1 BaseScope signals and immunohistochemical detection of YBX1 expression in NCE (n = 81), LSIL (n = 169), HSIL (n = 187) and CSCC (n = 253) tissues (*P < 0.05, **P < 0.01, ***P < 0.001, Kruskal–Wallis Test). C Box plot analysis on NCE, LSIL, HSIL and CSCC tissues (n = 690) demonstrated the significant Spearman correlation between circPOLD1 and YBX1 expressions (r = 0.497 (95% confidence interval = 0.4365 to 0.5524), p < 0.0001). D In identifying LSIL + (LSIL and worse), the ROC curves of circPOLD1 (area under curve, AUC = 0.7656, P < 0.0001) and circPOLD1/YBX1panel (AUC = 0.9514, P < 0.0001). E In identifying HSIL + (HSIL and worse), the ROC curves of circPOLD1 (AUC = 0.7282, P < 0.001) and circPOLD1/YBX1 panel (AUC = 0.8165, P < 0.0001). F The relative expression of cirPOLD1 in serum specimens of an independent validation set comprising 30 normal controls, 40 HSIL patients and 50 cervical cancer patients by qRT-PCR validation. G The diagnostic performance of HSIL diagnosis by circPOLD1 in the serum-based validation set via ROC analysis. (AUC = 0.731, P = 0.001). H The diagnostic performance of cervical cancer diagnosis by circPOLD1 in the serum-based validation set. (AUC = 0.858, P < 0.0001)

Journal: Journal of Translational Medicine

Article Title: The biomarker potential of circPOLD1 and its binding protein YBX1 in cervical carcinogenesis

doi: 10.1186/s12967-025-06494-3

Figure Lengend Snippet: The biomarker potential of circPOLD1 in cervical carcinogenesis. A Reprehensive images of BaseScope staining of circPOLD1 (red arrows) and immunohistochemical staining of YBX1 expression in NCE, LSIL, HSIL, and CSCC tissues. The scale bar represents 50 um. B Statistics analysis of density of circPOLD1 BaseScope signals and immunohistochemical detection of YBX1 expression in NCE (n = 81), LSIL (n = 169), HSIL (n = 187) and CSCC (n = 253) tissues (*P < 0.05, **P < 0.01, ***P < 0.001, Kruskal–Wallis Test). C Box plot analysis on NCE, LSIL, HSIL and CSCC tissues (n = 690) demonstrated the significant Spearman correlation between circPOLD1 and YBX1 expressions (r = 0.497 (95% confidence interval = 0.4365 to 0.5524), p < 0.0001). D In identifying LSIL + (LSIL and worse), the ROC curves of circPOLD1 (area under curve, AUC = 0.7656, P < 0.0001) and circPOLD1/YBX1panel (AUC = 0.9514, P < 0.0001). E In identifying HSIL + (HSIL and worse), the ROC curves of circPOLD1 (AUC = 0.7282, P < 0.001) and circPOLD1/YBX1 panel (AUC = 0.8165, P < 0.0001). F The relative expression of cirPOLD1 in serum specimens of an independent validation set comprising 30 normal controls, 40 HSIL patients and 50 cervical cancer patients by qRT-PCR validation. G The diagnostic performance of HSIL diagnosis by circPOLD1 in the serum-based validation set via ROC analysis. (AUC = 0.731, P = 0.001). H The diagnostic performance of cervical cancer diagnosis by circPOLD1 in the serum-based validation set. (AUC = 0.858, P < 0.0001)

Article Snippet: Small interference RNAs (siRNAs) that target the back-spliced junction of circPOLD1 were designed and synthesized by GenePharma (Shanghai, China).

Techniques: Biomarker Discovery, Staining, Immunohistochemical staining, Expressing, Quantitative RT-PCR, Diagnostic Assay

clinicopathological characteristics of patients with cervical cancer

Journal: Journal of Translational Medicine

Article Title: The biomarker potential of circPOLD1 and its binding protein YBX1 in cervical carcinogenesis

doi: 10.1186/s12967-025-06494-3

Figure Lengend Snippet: clinicopathological characteristics of patients with cervical cancer

Article Snippet: Small interference RNAs (siRNAs) that target the back-spliced junction of circPOLD1 were designed and synthesized by GenePharma (Shanghai, China).

Techniques: Expressing

Bioinformatics analysis and screening of differentially expressed genes between wild-type control and BL-I PF lung tissues of mice in the gene expression omnibus database. (a) The heat map and volcano plot of differentiating genes in the GSE43695 dataset. (b) The heat map and volcano plot of differentiating genes in the GSE123293 dataset. (c) The Venn diagram illustrating the intersection of low-expression differential genes in the GSE123293 and GSE43695 datasets. (d) The Venn diagram illustrating the intersection of high-expression differential genes in the GSE123293 and GSE43695 datasets. (e) Intersection of highly expressed distinguishing genes in the GSE123293 and GSE43695 datasets (difference multiple >2). Zbtb16: Zinc finger and BTB domain containing 16, HPGD: 15-hydroxyprostaglandin dehydrogenase, AOX3: Aldehyde oxidase 3, DBP: D-box binding PAR BZIP transcription factor, S100A8: S100 calcium binding protein A8, S100A9: S100 calcium binding protein A9, BL-I: Bleomycin-induced, PF: Pulmonary fibrosis.

Journal: CytoJournal

Article Title: Zinc finger and broad-complex, tramtrack, and bric-a-brac domain containing 16 silencing attenuates bleomycin-induced pulmonary fibrosis in mice through inhibition of the phosphoinositide 3-kinase/protein kinase B/mammalian target of rapamycin pathway

doi: 10.25259/Cytojournal_223_2024

Figure Lengend Snippet: Bioinformatics analysis and screening of differentially expressed genes between wild-type control and BL-I PF lung tissues of mice in the gene expression omnibus database. (a) The heat map and volcano plot of differentiating genes in the GSE43695 dataset. (b) The heat map and volcano plot of differentiating genes in the GSE123293 dataset. (c) The Venn diagram illustrating the intersection of low-expression differential genes in the GSE123293 and GSE43695 datasets. (d) The Venn diagram illustrating the intersection of high-expression differential genes in the GSE123293 and GSE43695 datasets. (e) Intersection of highly expressed distinguishing genes in the GSE123293 and GSE43695 datasets (difference multiple >2). Zbtb16: Zinc finger and BTB domain containing 16, HPGD: 15-hydroxyprostaglandin dehydrogenase, AOX3: Aldehyde oxidase 3, DBP: D-box binding PAR BZIP transcription factor, S100A8: S100 calcium binding protein A8, S100A9: S100 calcium binding protein A9, BL-I: Bleomycin-induced, PF: Pulmonary fibrosis.

Article Snippet: Both small interfere RNA (siRNAs) targeting Zbtb16 (si-Zbtb16) and its negative control (si-NC) were purchased from GenePharma (Shanghai, China).

Techniques: Control, Gene Expression, Expressing, Binding Assay

Zbtb16 silencing mitigated BL-I PF and inflammation in mice. (a and b) The Zbtb16 mRNA and protein expression levels in lung tissues of mice were measured via qRT-PCR and Western blot. n = 3; ✶ ✶ P < 0.01; ✶ ✶ ✶ P < 0.001. Zbtb16: Zinc finger and BTB domain containing 16, NC: Negative control, BL-I: Bleomycin-induced, RT-qPCR: Reverse transcription quantitative polymerase chain reaction, PF: Pulmonary fibrosis.

Journal: CytoJournal

Article Title: Zinc finger and broad-complex, tramtrack, and bric-a-brac domain containing 16 silencing attenuates bleomycin-induced pulmonary fibrosis in mice through inhibition of the phosphoinositide 3-kinase/protein kinase B/mammalian target of rapamycin pathway

doi: 10.25259/Cytojournal_223_2024

Figure Lengend Snippet: Zbtb16 silencing mitigated BL-I PF and inflammation in mice. (a and b) The Zbtb16 mRNA and protein expression levels in lung tissues of mice were measured via qRT-PCR and Western blot. n = 3; ✶ ✶ P < 0.01; ✶ ✶ ✶ P < 0.001. Zbtb16: Zinc finger and BTB domain containing 16, NC: Negative control, BL-I: Bleomycin-induced, RT-qPCR: Reverse transcription quantitative polymerase chain reaction, PF: Pulmonary fibrosis.

Article Snippet: Both small interfere RNA (siRNAs) targeting Zbtb16 (si-Zbtb16) and its negative control (si-NC) were purchased from GenePharma (Shanghai, China).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Negative Control, Reverse Transcription, Real-time Polymerase Chain Reaction

Zbtb16 silencing mitigated BL-I PF and inflammation in mice. (a) Lung tissue sections were stained with anti-Zbtb16 antibody by IHC to evaluate Zbtb16 protein expression. The red arrows indicate Zbtb16 protein-positive expression in this region of the tissue. (magnification: 100x) (b) Lung tissue sections were stained with H&E to examine alveolar structure and inflammatory cell infiltration. The red arrows indicate extensive inflammatory infiltration in this region of the tissue. (magnification: 100x) (c) Lung tissue sections were stained with Masson’s trichrome to evaluate collagen fiber deposition. The red arrows indicate significant collagen fiber deposition in this region of the tissue. (magnification: 100) (d) Lung tissue sections were subjected to TUNEL staining to assess apoptotic cell levels. The red arrows indicate a prominent aggregation of TUNEL-positive cells in this region of the tissue. (magnification: 100x) n = 5; ✶ ✶ P < 0.01; ✶ ✶ ✶ P < 0.001. Zbtb16: Zinc finger and BTB domain containing 16, NC: Negative control, IHC: Immunohistochemical, H&E: Hematoxylin and eosin, BL-I: Bleomycin-induced, TUNEL: Terminal deoxynucleotidyl transferase dUTP nick-end labeling, PF: Pulmonary fibrosis.

Journal: CytoJournal

Article Title: Zinc finger and broad-complex, tramtrack, and bric-a-brac domain containing 16 silencing attenuates bleomycin-induced pulmonary fibrosis in mice through inhibition of the phosphoinositide 3-kinase/protein kinase B/mammalian target of rapamycin pathway

doi: 10.25259/Cytojournal_223_2024

Figure Lengend Snippet: Zbtb16 silencing mitigated BL-I PF and inflammation in mice. (a) Lung tissue sections were stained with anti-Zbtb16 antibody by IHC to evaluate Zbtb16 protein expression. The red arrows indicate Zbtb16 protein-positive expression in this region of the tissue. (magnification: 100x) (b) Lung tissue sections were stained with H&E to examine alveolar structure and inflammatory cell infiltration. The red arrows indicate extensive inflammatory infiltration in this region of the tissue. (magnification: 100x) (c) Lung tissue sections were stained with Masson’s trichrome to evaluate collagen fiber deposition. The red arrows indicate significant collagen fiber deposition in this region of the tissue. (magnification: 100) (d) Lung tissue sections were subjected to TUNEL staining to assess apoptotic cell levels. The red arrows indicate a prominent aggregation of TUNEL-positive cells in this region of the tissue. (magnification: 100x) n = 5; ✶ ✶ P < 0.01; ✶ ✶ ✶ P < 0.001. Zbtb16: Zinc finger and BTB domain containing 16, NC: Negative control, IHC: Immunohistochemical, H&E: Hematoxylin and eosin, BL-I: Bleomycin-induced, TUNEL: Terminal deoxynucleotidyl transferase dUTP nick-end labeling, PF: Pulmonary fibrosis.

Article Snippet: Both small interfere RNA (siRNAs) targeting Zbtb16 (si-Zbtb16) and its negative control (si-NC) were purchased from GenePharma (Shanghai, China).

Techniques: Staining, Expressing, TUNEL Assay, Negative Control, Immunohistochemical staining

Zbtb16 silencing mitigated BL-I PF and inflammation in mice. (a) The wet/dry ratio of lung tissue was measured. (b) The albumin concentration in BLF of mice lung was assessed by ELISA. (c and d) The protein levels of collagen I, Fn, and α-SMA in lung tissues of mice were assessed by Western blot. (e) The levels of inflammatory factors TNF-α, IL-6, and IL-1β in BLF of mice lung were assessed by ELISA. n = 3; ✶ P < 0.05; ✶ ✶ P < 0.01; ✶ ✶ ✶ P < 0.001. Zbtb16: Zinc finger and BTB domain containing 16, NC: Negative control, TNF-α: Tumor necrosis factor-α, IL-6: Interleukin 6, IL-1β: interleukin 1β, Fn: Fibronectin, α-SMA: α-smooth muscle actin, ALB: Albumin; ELISA: Enzyme-linked immunosorbent assay, Fn: Fibronectin, BLF: Bronchoalveolar lavage fluid, PF: Pulmonary fibrosis.

Journal: CytoJournal

Article Title: Zinc finger and broad-complex, tramtrack, and bric-a-brac domain containing 16 silencing attenuates bleomycin-induced pulmonary fibrosis in mice through inhibition of the phosphoinositide 3-kinase/protein kinase B/mammalian target of rapamycin pathway

doi: 10.25259/Cytojournal_223_2024

Figure Lengend Snippet: Zbtb16 silencing mitigated BL-I PF and inflammation in mice. (a) The wet/dry ratio of lung tissue was measured. (b) The albumin concentration in BLF of mice lung was assessed by ELISA. (c and d) The protein levels of collagen I, Fn, and α-SMA in lung tissues of mice were assessed by Western blot. (e) The levels of inflammatory factors TNF-α, IL-6, and IL-1β in BLF of mice lung were assessed by ELISA. n = 3; ✶ P < 0.05; ✶ ✶ P < 0.01; ✶ ✶ ✶ P < 0.001. Zbtb16: Zinc finger and BTB domain containing 16, NC: Negative control, TNF-α: Tumor necrosis factor-α, IL-6: Interleukin 6, IL-1β: interleukin 1β, Fn: Fibronectin, α-SMA: α-smooth muscle actin, ALB: Albumin; ELISA: Enzyme-linked immunosorbent assay, Fn: Fibronectin, BLF: Bronchoalveolar lavage fluid, PF: Pulmonary fibrosis.

Article Snippet: Both small interfere RNA (siRNAs) targeting Zbtb16 (si-Zbtb16) and its negative control (si-NC) were purchased from GenePharma (Shanghai, China).

Techniques: Concentration Assay, Enzyme-linked Immunosorbent Assay, Western Blot, Negative Control

Zbtb16 silencing alleviated T-β1-I mice LEC fibroblasts and inflammation. (a and b) The Zbtb16 mRNA and protein expression levels in mice LECs were measured by qRT-PCR and Western blot. (c and d) The protein levels of collagen I, Fn, and α-SMA in mice LECs were assessed by Western blot. (e) The levels of inflammatory factors TNF-α, IL-6, and IL-1β in cell culture medium supernatant were assessed by ELISA. n = 3; ✶ P < 0.05; ✶ ✶ P < 0.01; ✶ ✶ ✶ P < 0.001. Zbtb16: Zinc finger and BTB domain containing 16, NC: Negative control, TNF-α: Tumor necrosis factor-α, IL-6: Interleukin 6, IL-1β: Interleukin 1β, Fn: Fibronectin, α-SMA: α-smooth muscle actin, ALB: Albumin, ELISA: Enzyme-linked immunosorbent assay, Fn: Fibronectin, LEC: Lung epithelial cell.

Journal: CytoJournal

Article Title: Zinc finger and broad-complex, tramtrack, and bric-a-brac domain containing 16 silencing attenuates bleomycin-induced pulmonary fibrosis in mice through inhibition of the phosphoinositide 3-kinase/protein kinase B/mammalian target of rapamycin pathway

doi: 10.25259/Cytojournal_223_2024

Figure Lengend Snippet: Zbtb16 silencing alleviated T-β1-I mice LEC fibroblasts and inflammation. (a and b) The Zbtb16 mRNA and protein expression levels in mice LECs were measured by qRT-PCR and Western blot. (c and d) The protein levels of collagen I, Fn, and α-SMA in mice LECs were assessed by Western blot. (e) The levels of inflammatory factors TNF-α, IL-6, and IL-1β in cell culture medium supernatant were assessed by ELISA. n = 3; ✶ P < 0.05; ✶ ✶ P < 0.01; ✶ ✶ ✶ P < 0.001. Zbtb16: Zinc finger and BTB domain containing 16, NC: Negative control, TNF-α: Tumor necrosis factor-α, IL-6: Interleukin 6, IL-1β: Interleukin 1β, Fn: Fibronectin, α-SMA: α-smooth muscle actin, ALB: Albumin, ELISA: Enzyme-linked immunosorbent assay, Fn: Fibronectin, LEC: Lung epithelial cell.

Article Snippet: Both small interfere RNA (siRNAs) targeting Zbtb16 (si-Zbtb16) and its negative control (si-NC) were purchased from GenePharma (Shanghai, China).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Cell Culture, Enzyme-linked Immunosorbent Assay, Negative Control

Bioinformatics predicted that Zbtb16 might be involved in BL-I PF of mice. (a) KEGG analysis predicted that Zbtb16 might be involved. (b) GESA analysis predicted that PAmT-P of Zbtb16 might be involved. (c) CO-IP was used to analyze the combination of AKT and Zbtb16. (d) The PAmT-P-related protein expression levels in mice lung tissues and mice LECs were measured by Western blot. (e) p-AKT/AKT and p-mTOR/mTOR levels were shown in BL-I PF lung tissues of mice; (f) p-AKT/AKT and p-mTOR/mTOR levels were shown in T-β1-I fibrosis mice LECs. n = 3; ✶ P < 0.05; ✶ ✶ P < 0.01, ✶ ✶ ✶ P < 0.001. Zbtb16: Zinc finger and BTB domain containing 16, NC: Negative control, AKT: Protein kinase B, p-AKT: Phosphorylated AKT, mTOR: Mammalian target of rapamycin, p-mTOR: Phosphorylated mTOR, TNF-α: Tumor necrosis factor-α, IL-6: Interleukin 6, IL-1β: Interleukin 1β, Fn: Fibronectin, α-SMA: α-smooth muscle actin, ALB: Albumin, ELISA: Enzyme-linked immunosorbent assay, LEC: Lung epithelial cell, BL-I: Bleomycin-induced, GESA: Gene set enrichment analysis, KEGG: Kyoto encyclopedia of genes and genomes, PAmT-P: PI3K/AKT/mTOR pathway, PF: Pulmonary fibrosis.

Journal: CytoJournal

Article Title: Zinc finger and broad-complex, tramtrack, and bric-a-brac domain containing 16 silencing attenuates bleomycin-induced pulmonary fibrosis in mice through inhibition of the phosphoinositide 3-kinase/protein kinase B/mammalian target of rapamycin pathway

doi: 10.25259/Cytojournal_223_2024

Figure Lengend Snippet: Bioinformatics predicted that Zbtb16 might be involved in BL-I PF of mice. (a) KEGG analysis predicted that Zbtb16 might be involved. (b) GESA analysis predicted that PAmT-P of Zbtb16 might be involved. (c) CO-IP was used to analyze the combination of AKT and Zbtb16. (d) The PAmT-P-related protein expression levels in mice lung tissues and mice LECs were measured by Western blot. (e) p-AKT/AKT and p-mTOR/mTOR levels were shown in BL-I PF lung tissues of mice; (f) p-AKT/AKT and p-mTOR/mTOR levels were shown in T-β1-I fibrosis mice LECs. n = 3; ✶ P < 0.05; ✶ ✶ P < 0.01, ✶ ✶ ✶ P < 0.001. Zbtb16: Zinc finger and BTB domain containing 16, NC: Negative control, AKT: Protein kinase B, p-AKT: Phosphorylated AKT, mTOR: Mammalian target of rapamycin, p-mTOR: Phosphorylated mTOR, TNF-α: Tumor necrosis factor-α, IL-6: Interleukin 6, IL-1β: Interleukin 1β, Fn: Fibronectin, α-SMA: α-smooth muscle actin, ALB: Albumin, ELISA: Enzyme-linked immunosorbent assay, LEC: Lung epithelial cell, BL-I: Bleomycin-induced, GESA: Gene set enrichment analysis, KEGG: Kyoto encyclopedia of genes and genomes, PAmT-P: PI3K/AKT/mTOR pathway, PF: Pulmonary fibrosis.

Article Snippet: Both small interfere RNA (siRNAs) targeting Zbtb16 (si-Zbtb16) and its negative control (si-NC) were purchased from GenePharma (Shanghai, China).

Techniques: Co-Immunoprecipitation Assay, Expressing, Western Blot, Negative Control, Enzyme-linked Immunosorbent Assay

Zbtb16 silencing alleviated T-β1-I mice LEC fibrosis by blocking PAmT-P. (a and b) The PAmT-P-related protein and collagen I, Fn, and α-SMA protein expression levels in mice LECs were measured through Western blot. (c) The TNF-α, IL-6, and IL-1β concentrations in mice LEC culture medium supernatant were detected by ELISA. n = 3; ✶ P < 0.05; ✶ ✶ P < 0.01; ✶ ✶ ✶ P < 0.001. Zbtb16: Zinc finger and BTB domain containing 16, NC: Negative control, AKT: Protein kinase B, p-AKT: Phosphorylated AKT, mTOR: Mammalian target of rapamycin, p-mTOR: Phosphorylated mTOR, PAmT-P: PI3K/AKT/mTOR pathway, Fn: Fibronectin, α-SMA: α-smooth muscle actin, LEC: Lung epithelial cell, TNF-α: Tumor necrosis factor-α, IL-6: Interleukin 6, IL-1β: Interleukin 1β, ELISA: Enzyme-linked immunosorbent assay.

Journal: CytoJournal

Article Title: Zinc finger and broad-complex, tramtrack, and bric-a-brac domain containing 16 silencing attenuates bleomycin-induced pulmonary fibrosis in mice through inhibition of the phosphoinositide 3-kinase/protein kinase B/mammalian target of rapamycin pathway

doi: 10.25259/Cytojournal_223_2024

Figure Lengend Snippet: Zbtb16 silencing alleviated T-β1-I mice LEC fibrosis by blocking PAmT-P. (a and b) The PAmT-P-related protein and collagen I, Fn, and α-SMA protein expression levels in mice LECs were measured through Western blot. (c) The TNF-α, IL-6, and IL-1β concentrations in mice LEC culture medium supernatant were detected by ELISA. n = 3; ✶ P < 0.05; ✶ ✶ P < 0.01; ✶ ✶ ✶ P < 0.001. Zbtb16: Zinc finger and BTB domain containing 16, NC: Negative control, AKT: Protein kinase B, p-AKT: Phosphorylated AKT, mTOR: Mammalian target of rapamycin, p-mTOR: Phosphorylated mTOR, PAmT-P: PI3K/AKT/mTOR pathway, Fn: Fibronectin, α-SMA: α-smooth muscle actin, LEC: Lung epithelial cell, TNF-α: Tumor necrosis factor-α, IL-6: Interleukin 6, IL-1β: Interleukin 1β, ELISA: Enzyme-linked immunosorbent assay.

Article Snippet: Both small interfere RNA (siRNAs) targeting Zbtb16 (si-Zbtb16) and its negative control (si-NC) were purchased from GenePharma (Shanghai, China).

Techniques: Blocking Assay, Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Negative Control