Journal:

Article Title: Assembly of cyclin D-dependent kinase and titration of p27 Kip1 regulated by mitogen-activated protein kinase kinase (MEK1)

doi:

Figure Lengend Snippet: Molecular correlates of S phase entry. (A) Cells from five individual subclones expressing inducible MEK1* were serum-starved and restimulated with ZnSO4 or FBS for 16 hr, and the S phase fraction was determined by using flow cytometry to estimate the DNA content of propidium iodide-stained nuclei. (B) Serum-starved cells (subclone 3) stimulated with ZnSO4 or FBS were assayed at 4-hr intervals for DNA content (Upper). Cyclin A, p21Cip1, and p27Kip1 levels were determined by immunoblotting. Precipitates recovered with antibodies to cyclin E and CDK2 were assayed for histone H1 kinase activity.

Article Snippet: Rabbit polyclonal antibodies against ERK1 (K-23), ERK2 (C-14), cyclin E (M-20), cyclin A (C-19), and p21 Cip1 (C-19) were purchased from Santa Cruz Biotechnology.

Techniques: Expressing, Flow Cytometry, Staining, Western Blot, Activity Assay

Journal:

Article Title: Assembly of cyclin D-dependent kinase and titration of p27 Kip1 regulated by mitogen-activated protein kinase kinase (MEK1)

doi:

Figure Lengend Snippet: Overexpression of cyclin D1, CDK4, and MEK1* induces cyclin A synthesis and CDK2 activation, but not p27Kip1 degradation. Cells (subclone 2) ectopically expressing cyclin D1, CDK4, and inducible MEK1* were starved and restimulated with ZnSO4 or FBS. Cells harvested at intervals thereafter (hr) were assayed for cyclin A and p27Kip1 expression by immunoblotting (top two panels), for cyclin D1-associated p27Kip1 by immunoprecipitation with cyclin D1 followed by immunoblotting with anti-p27Kip1 (middle panel), and for cyclin E- and CDK2-associated histone H1 kinase activity (bottom two panels). More than 85% of p27Kip1 coprecipitated with cyclin D1.

Article Snippet: Rabbit polyclonal antibodies against ERK1 (K-23), ERK2 (C-14), cyclin E (M-20), cyclin A (C-19), and p21 Cip1 (C-19) were purchased from Santa Cruz Biotechnology.

Techniques: Over Expression, Activation Assay, Expressing, Western Blot, Immunoprecipitation, Activity Assay

Journal: Journal of Biological Chemistry

Article Title: Selective Enhancing Effect of Early Mitotic Inhibitor 1 (Emi1) Depletion on the Sensitivity of Doxorubicin or X-ray Treatment in Human Cancer Cells

doi: 10.1074/jbc.m112.446351

Figure Lengend Snippet: FIGURE 7. Synergistic anticancer effect of Emi1 knockdown in combination with doxorubucin in normal cells. A, Emi1 or control siRNA was transfected into NHDF cells, and cells were collected after 48 h. The indicated proteins in siRNA-treated NHDF cells were examined by Western blotting. B, Emi1 siRNA and/or cyclin A siRNA were transfected into NHDF cells. The left panel shows the expression of Emi1 and cyclin A examined by Western blot analysis after 48 h of siRNA transfection. -Actin expression was used as a loading control. The right panel shows DAPI staining and percentage of cells with polyploidy after Emi1 siRNA and/or cyclin A siRNA transfection. Cells were stained with DAPI to visualize the nuclei, and percentage of cells with polyploidy was determined by DNA content analysis after PI staining using a flow cytometer. *, p 0.05. C, flow cytometric analysis of annexin V and PI staining in control and Emi1 siRNA treated NHDF or HFL III cells after treatment with doxorubucin (DOXY; 0.5 g/ml) for 12 h. We performed three independent experiments.

Article Snippet: Human cyclin A siRNA (sc-29282) and human E2F1 siRNA (sc-61861) were obtained from Santa Cruz Biotechnology.

Techniques: Knockdown, Control, Transfection, Western Blot, Expressing, Staining, Flow Cytometry

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Super enhancer-driven LINC01013 mediates hypoxia-induced mitochondrial dysfunction by HSPA9 to determine pulmonary arterial smooth muscle cell fate

doi: 10.1007/s00018-025-06071-3

Figure Lengend Snippet: Silencing of LINC01013 inhibited hypoxia-induced hPASMC proliferation and inflammation. A , B hPASMC proliferation was determined by CCK8 and EdU incorporation assays ( n = 6). EdU (red), DAPI (blue), scale bar, 50 μm. C Western blotting analysis of PCNA, cyclin A and cyclin D in hPASMCs ( n = 6). D RT‒qPCR analysis showed the mRNA levels of TNF-α and IL-6 after LINC01013 knockdown, β-actin was used as a internal reference gene ( n = 6). E Western blotting analysis of TNF-α and IL-6 in hPASMCs. ( n = 6). All values are presented as the mean ± SEM. Statistical analysis was performed with one-way ANOVA. * p < 0.05, ** p < 0.01, *** p < 0.001. Nor, normoxia; Hyp, hypoxia; NC, negative control; si, siRNA

Article Snippet: The antibody against CEBPB (PB9171, BA0670, 1:500, Boster, Wuhan, China), H3K27ac (A7253, 1:500, ABclonal, Wuhan, China), H3K4me1 (A2355, 1:500, Wuhan, China), PCNA (A00125, 1:500, Boster, Wuhan, China), Cyclin A (PB0515, 1:500, Boster, Wuhan, China), Cyclin D (BM4272, 1:500, Boster, Wuhan, China), IL-6 (AF7236, 1:500, Beyotime, Shanghai, China), TNF-α (AF8208, 1:500, Beyotime, Shanghai, China), PKM2 (4053, 1:1000, Cell Signaling, MA, US), HK II (66974-1-Ig, 1:1000, Proteintech, IL, USA), PDH (2784, 1:1000, Cell Signaling, MA, US), HSPA9 (14887-1-AP, 1:5000, Proteintech, IL, USA), VDAC1 (10866-1-AP, 1:5000, Proteintech, IL, USA), and β-actin (TA-09, 1:1000, ZSGB‐BIO, Beijing, China) was incubated at 4 °C overnight, followed by incubation with appropriate horseradish peroxidase-conjugated secondary antibodies at room temperature for 1 h, and proteins were visualized with enhanced chemiluminescence reagents.

Techniques: Western Blot, Knockdown, Negative Control

Journal: Frontiers in Immunology

Article Title: Multi-omics analysis and experimental validation of the value of monocyte-associated features in prostate cancer prognosis and immunotherapy

doi: 10.3389/fimmu.2024.1426474

Figure Lengend Snippet: CCNA2 identified as the best prognostic gene among monocyte-associated genes. (A, B) Functional analysis of monocyte-related prognostic genes. (C, D) GBM and RandomForest algorithms to screen key prognostic genes in the TCGA-PRAD dataset. (E, F) GBM and RandomForest algorithms to screen key prognostic genes in the GSE16560 dataset. (G, H) KM curves of CCNA2 and ACSM3 in the TCGA-PRAD dataset. (I, J) KM curves of CCNA2 and ACSM3 in the GSE16560 dataset.

Article Snippet: Next, CCNA2 antibody (BOSTER, PB9424) was applied dropwise to cover the tissue chip, which was left at room temperature for 2 hours.

Techniques: Functional Assay

Journal: Frontiers in Immunology

Article Title: Multi-omics analysis and experimental validation of the value of monocyte-associated features in prostate cancer prognosis and immunotherapy

doi: 10.3389/fimmu.2024.1426474

Figure Lengend Snippet: CCNA2 positively correlates with monocyte infiltration levels. (A) Correlation analysis of CCNA2 and monocyte infiltration levels. (B, C) Analysis of CCNA2 correlation with monocyte markers. (D) Mendelian randomization analysis of high HLA-DR expressing monocytes in relation to prostate cancer. (E–I) Single-cell analysis of the correlation between CCNA2 and immune cell infiltration.

Article Snippet: Next, CCNA2 antibody (BOSTER, PB9424) was applied dropwise to cover the tissue chip, which was left at room temperature for 2 hours.

Techniques: Expressing, Single-cell Analysis

Journal: Frontiers in Immunology

Article Title: Multi-omics analysis and experimental validation of the value of monocyte-associated features in prostate cancer prognosis and immunotherapy

doi: 10.3389/fimmu.2024.1426474

Figure Lengend Snippet: Functional analysis of CCNA2 in PRAD. (A) KEGG analysis of CCNA2 in PRAD. (B–J) GSEA analysis of CCNA2 in PRAD.

Article Snippet: Next, CCNA2 antibody (BOSTER, PB9424) was applied dropwise to cover the tissue chip, which was left at room temperature for 2 hours.

Techniques: Functional Assay

Journal: Frontiers in Immunology

Article Title: Multi-omics analysis and experimental validation of the value of monocyte-associated features in prostate cancer prognosis and immunotherapy

doi: 10.3389/fimmu.2024.1426474

Figure Lengend Snippet: CCNA2 has a high binding capacity to PRAD-targeted drugs. (A) Analysis of the binding capacity of CCNA2 to PD1 inhibitors. (B) Analysis of the binding capacity of CCNA2 to bicalutamide. (C) Analysis of the binding capacity of CCNA2 to enzalutamide. (D) Analysis of the binding capacity of CCNA2 to abiraterone.

Article Snippet: Next, CCNA2 antibody (BOSTER, PB9424) was applied dropwise to cover the tissue chip, which was left at room temperature for 2 hours.

Techniques: Binding Assay

Journal: Frontiers in Immunology

Article Title: Multi-omics analysis and experimental validation of the value of monocyte-associated features in prostate cancer prognosis and immunotherapy

doi: 10.3389/fimmu.2024.1426474

Figure Lengend Snippet: CCNA2 is highly expressed in PRAD and is associated with poor patient prognosis. (A, B) Differential expression of CCNA2 in PRAD. (C) Diagnostic predictive value of CCNA2 in PRAD. (D) KM curve of overall survival of CCNA2 in PRAD. (E) Prognostic predictive value of CCNA2 in PRAD.

Article Snippet: Next, CCNA2 antibody (BOSTER, PB9424) was applied dropwise to cover the tissue chip, which was left at room temperature for 2 hours.

Techniques: Quantitative Proteomics, Diagnostic Assay

Journal: Molecular Vision

Article Title: Small interfering RNA targeting of S phase kinase-interacting protein 2 inhibits cell proliferation of pterygium fibroblasts

doi:

Figure Lengend Snippet: Down-regulation of Skp2 protein by siRNA in PFC in vitro by immunoflurescence. Immunoflurescence staining indicated high constitutive levels of Skp2 protein (arrow indication) in the nucleolus of PFC and NFC transfected with pSuppressor vehicle ( A , D ) or without transfection ( B , E ). Transfection with Skp2 siRNA dramatically decreased the expression of Skp2 protein in PFC cells ( C , F ). The scale bar is equal to 40 μm.

Article Snippet: Skp2 antibody and the Steptavidin-Alkaline Phosphatase Complex (SABC) kit were purchased from the Boster Company (Wuhan, China).

Techniques: In Vitro, Staining, Transfection, Expressing

Journal: Molecular Vision

Article Title: Small interfering RNA targeting of S phase kinase-interacting protein 2 inhibits cell proliferation of pterygium fibroblasts

doi:

Figure Lengend Snippet: Down-regulation of Skp2 protein by siRNA in PFC in vivo by immunoflurescence. Immunoflurescence staining indicated high constitutive levels of Skp2 protein in the nucleolus of PFC and NFC transfected with pSuppressor vehicle ( A , D ) or without transfection ( B , E ). Transfection with Skp2 siRNA dramatically decreased the expression of Skp2 protein in PFC cells ( C , F ). The scale bar is equal to 40 μm.

Article Snippet: Skp2 antibody and the Steptavidin-Alkaline Phosphatase Complex (SABC) kit were purchased from the Boster Company (Wuhan, China).

Techniques: In Vivo, Staining, Transfection, Expressing

Journal: Molecular Vision

Article Title: Small interfering RNA targeting of S phase kinase-interacting protein 2 inhibits cell proliferation of pterygium fibroblasts

doi:

Figure Lengend Snippet: Skp2 siRNA induced p27 kip1 accumulationin in PFC in vitro by immunofluorescence. Immunofluorescence staining indicated the expression of p27 kip1 dramatically increased in PFC and NFC transfection with Skp2 siRNA ( C , F ) when compared with PFC transfection with pSuppressor vehicle ( A , D ) or without transfection ( B , E ). The scale bar is equal to 40 μm.

Article Snippet: Skp2 antibody and the Steptavidin-Alkaline Phosphatase Complex (SABC) kit were purchased from the Boster Company (Wuhan, China).

Techniques: In Vitro, Immunofluorescence, Staining, Expressing, Transfection

Journal: Molecular Vision

Article Title: Small interfering RNA targeting of S phase kinase-interacting protein 2 inhibits cell proliferation of pterygium fibroblasts

doi:

Figure Lengend Snippet: Skp2 siRNA induced p27 kip1 accumulationin in PFC in vivo by immunofluorescence. Immunofluorescence staining indicated the expression of p27 kip1 dramatically increased in PFC and NFC transfection with Skp2 siRNA ( C , F ) when compared with PFC transfection with pSuppressor vehicle ( A , D ) or without transfection ( B , E ). The scale bar is equal to 40 μm.

Article Snippet: Skp2 antibody and the Steptavidin-Alkaline Phosphatase Complex (SABC) kit were purchased from the Boster Company (Wuhan, China).

Techniques: In Vivo, Immunofluorescence, Staining, Expressing, Transfection

Journal: Molecular Vision

Article Title: Small interfering RNA targeting of S phase kinase-interacting protein 2 inhibits cell proliferation of pterygium fibroblasts

doi:

Figure Lengend Snippet: Cell viability assay by MTT. After 6 to 12 days transfection with Skp2 siRNA, cell viability was significantly decreased in cultured hPFC cells compared with vehicle control and blank control epsecially on the 6th day after transfection. *p<0.01 versus vehicle and blank control; **p<0.05 versus vehicle and blank control.

Article Snippet: Skp2 antibody and the Steptavidin-Alkaline Phosphatase Complex (SABC) kit were purchased from the Boster Company (Wuhan, China).

Techniques: Viability Assay, Transfection, Cell Culture, Control

Journal: Molecular Vision

Article Title: Small interfering RNA targeting of S phase kinase-interacting protein 2 inhibits cell proliferation of pterygium fibroblasts

doi:

Figure Lengend Snippet: Skp2 siRNA inhibited the cell proliferation of PFC in vitro (Brdu). For Brdu incorporation, cells growing on coverslips were incubated with Brdu. Incorporated Brdu was detected with antibodies as described in the Methods. Statistical analysis after cell counting showed that Brdu positive cells decreased after hPFC transfection with Skp2 siRNA compared with control cells. **p<0.01 versus vehicle and blank control.

Article Snippet: Skp2 antibody and the Steptavidin-Alkaline Phosphatase Complex (SABC) kit were purchased from the Boster Company (Wuhan, China).

Techniques: In Vitro, BrdU Incorporation Assay, Incubation, Cell Counting, Transfection, Control

Journal: Molecular Vision

Article Title: Small interfering RNA targeting of S phase kinase-interacting protein 2 inhibits cell proliferation of pterygium fibroblasts

doi:

Figure Lengend Snippet: Skp2 siRNA inhibited the cell proliferation of PFC in vivo (Brdu). For Brdu incorporation, ptergium tissue were incubated with Brdu. Incorporated Brdu was detected with antibodies as described in the Methods. Statistical analysis after cell counting showed that Brdu positive cells decreased after hPFC transfection with Skp2 siRNA compared with control cells. **p<0.01 versus vehicle and blank control.

Article Snippet: Skp2 antibody and the Steptavidin-Alkaline Phosphatase Complex (SABC) kit were purchased from the Boster Company (Wuhan, China).

Techniques: In Vivo, BrdU Incorporation Assay, Incubation, Cell Counting, Transfection, Control

Journal: Molecular Vision

Article Title: Small interfering RNA targeting of S phase kinase-interacting protein 2 inhibits cell proliferation of pterygium fibroblasts

doi:

Figure Lengend Snippet: Down-regulation of PCNA protein by siRNA in PFC in vitro and in vivo. Immunofluorescence staining indicated the expression of PCNA decreased in PFC and NFC transfection with Skp2 siRNA ( C , F ) when compared with hPFC and hNFC transfection with pSuppressor vehicle ( A , D ) or without transfection ( B , E ). The scale bar is equal to 40 μm.

Article Snippet: Skp2 antibody and the Steptavidin-Alkaline Phosphatase Complex (SABC) kit were purchased from the Boster Company (Wuhan, China).

Techniques: In Vitro, In Vivo, Immunofluorescence, Staining, Expressing, Transfection

Journal: Molecular Vision

Article Title: Small interfering RNA targeting of S phase kinase-interacting protein 2 inhibits cell proliferation of pterygium fibroblasts

doi:

Figure Lengend Snippet: Down-regulation of PCNA protein by siRNA in PFC in vitro and in vivo. Immunofluorescence staining indicated the expression of PCNA decreased in PFC and NFC transfection with Skp2 siRNA ( C , F ) when compared with hPFC and hNFC transfection with pSuppressor vehicle ( A , D ) or without transfection ( B , E ). The scale bar is equal to 40 μm.

Article Snippet: Skp2 antibody and the Steptavidin-Alkaline Phosphatase Complex (SABC) kit were purchased from the Boster Company (Wuhan, China).

Techniques: In Vitro, In Vivo, Immunofluorescence, Staining, Expressing, Transfection

Journal: Molecular Vision

Article Title: Small interfering RNA targeting of S phase kinase-interacting protein 2 inhibits cell proliferation of pterygium fibroblasts

doi:

Figure Lengend Snippet: Skp2 siRNA inhibited the proliferation of PFC and NFC in vivo. Hematoxylin and eosin staining was performed to examine the histological changes 14 days after transfection. Obvious PFC and NFC proliferation was detected in pSuppressor vehicle group or without transfection ( A , D , B , E ). There was little PFC and NFC proliferation in Skp2 siRNA transfection group ( C , F ). Scale bar is equal to 20 μm.

Article Snippet: Skp2 antibody and the Steptavidin-Alkaline Phosphatase Complex (SABC) kit were purchased from the Boster Company (Wuhan, China).

Techniques: In Vivo, Staining, Transfection