Journal: Cancer medicine

Article Title: Nuclear receptor coactivator 6 (NCoA6) promotes cell proliferation, migration, and invasion in pancreatic cancer.

doi: 10.1002/cam4.6427

Figure Lengend Snippet: FIGURE 3 NCoA6 knockdown decreases the proliferation, cell cycle, and apoptosis of pancreatic cancer cells. CCK-8 assays (A, B) and colony formation assays (C, D) were used to test the proliferation of PDAC cells transfected with NCoA6 shRNAs. The representative images and statistical results of the cell cycle (E, F) and apoptosis (G, H) using flow cytometry. (I, J) Western blotting analysis of p21 and p27 expression in NCoA6-silenced PANC-1 and SW1990 cells. (K, L) Western blotting analysis of CDK4, CDK2, Cyclin D1, Cyclin E1, and Cyclin A2 expression in NCoA6-silenced PANC-1 and SW1990 cells.

Article Snippet: Protein extraction and western blotting were performed as described in our previous research.16 Antibodies against NCoA6 (1:1000), E- cadherin (1:5000), N- cadherin (1:3000), FBW7 (1:1000), CDX2 (1:1000), cyclin- dependent kinase 4 (CDK4, 1:2000), cyclin- dependent kinase 2 (CDK2, 1:5000), Cyclin D1 (1:5000), Cyclin E1 (1:1000), and Cyclin A2 (1:2000) were obtained from Proteintech.

Techniques: Knockdown, CCK-8 Assay, Transfection, Flow Cytometry, Western Blot, Expressing

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 Bioengineering and Biotechnology

Article Title: MiR-29c-3p Suppresses the Migration, Invasion and Cell Cycle in Esophageal Carcinoma via CCNA2/p53 Axis

doi: 10.3389/fbioe.2020.00075

Figure Lengend Snippet: Primer sequence.

Article Snippet: In order to investigate the underlying mechanism of CCNA2 on EC cells, cells were classified into four groups: si-NC + DMSO, si-CCNA2 + DMSO, si-NC + PFTβand si-CCNA2 + PFTβ groups [PFTβ, p53 inhibitor, HY-16702, MedChemExpress, 10 μM ( )]. qRT-PCR was firstly performed to test CCNA2 level in each group, finding that CCNA2 was markedly decreased in cells transfected with si-CCNA2 + DMSO ( ).

Techniques: Sequencing

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: MiR-29c-3p Suppresses the Migration, Invasion and Cell Cycle in Esophageal Carcinoma via CCNA2/p53 Axis

doi: 10.3389/fbioe.2020.00075

Figure Lengend Snippet: MiR-29c-3p is decreased in EC tissues accompanied by low survival rate and associated with the increase of CCNA2. TCGA database was utilized to access expression data of miRNAs and mRNAs of ESCA, and (A) the results of differential analysis were plotted in Volcano plots, with red representing high expression and green representing low expression. In panel (B) , miR-29c-3p level in EC tissues were determined as shown in a box plot. (C) Survival analysis of miR-29c-3p in TCGA-ESCA dataset was performed, with the red line as high expression and blue line as low expression. In panel (D) , Venn diagram was made to find the candidate targets of miR-29c-3p, acquiring 10 DEmRNAs. In panel (E) , correlation analysis was conducted between miR-29c-3p and CCNA2 (–0.57) as plotted in a heat map. In panel (F) , CCNA2 expression in EC cells was examined. Clinical tissue samples were used to further explore the (G) expression of miR-29c-3p and CCNA2 mRNA in EC tissues by qRT-PCR, (H) the protein level of CCNA2 (P1, P2, P3 referred to three EC samples) via Western blot and (I) the correlation between miR-29c-3p and CCNA2. * P < 0.05.

Article Snippet: In order to investigate the underlying mechanism of CCNA2 on EC cells, cells were classified into four groups: si-NC + DMSO, si-CCNA2 + DMSO, si-NC + PFTβand si-CCNA2 + PFTβ groups [PFTβ, p53 inhibitor, HY-16702, MedChemExpress, 10 μM ( )]. qRT-PCR was firstly performed to test CCNA2 level in each group, finding that CCNA2 was markedly decreased in cells transfected with si-CCNA2 + DMSO ( ).

Techniques: Expressing, Quantitative RT-PCR, Western Blot

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: MiR-29c-3p Suppresses the Migration, Invasion and Cell Cycle in Esophageal Carcinoma via CCNA2/p53 Axis

doi: 10.3389/fbioe.2020.00075

Figure Lengend Snippet: MiR-29c-3p targets CCNA2 and inhibits its expression. Targeted binding sites of miR-29c-3p and CCNA2 were predicted before as shown in panel (A) . To investigate their targeted relationship, (B) dual-luciferase assay was performed to confirm their targeted binding, and (C) RIP was conducted to describe the effect of miR-29c-3p on CCNA2. Moreover, (D,E) qRT-PCR and Western blot were carried out to determine CCNA2 expression in mRNA and protein levels in miR-29c-3p mimic transfected cells, so as to further verify such relationship. * P < 0.05.

Article Snippet: In order to investigate the underlying mechanism of CCNA2 on EC cells, cells were classified into four groups: si-NC + DMSO, si-CCNA2 + DMSO, si-NC + PFTβand si-CCNA2 + PFTβ groups [PFTβ, p53 inhibitor, HY-16702, MedChemExpress, 10 μM ( )]. qRT-PCR was firstly performed to test CCNA2 level in each group, finding that CCNA2 was markedly decreased in cells transfected with si-CCNA2 + DMSO ( ).

Techniques: Expressing, Binding Assay, Luciferase, Quantitative RT-PCR, Western Blot, Transfection

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: MiR-29c-3p Suppresses the Migration, Invasion and Cell Cycle in Esophageal Carcinoma via CCNA2/p53 Axis

doi: 10.3389/fbioe.2020.00075

Figure Lengend Snippet: CCNA2 silencing regulates the migration, invasion and cell cycle in EC by promoting p53 signaling pathway. si-NC + DMSO, si-CCNA2 + DMSO, si-NC + PFTβ and si-CCNA2 + PFTβ were transfected into cells. qRT-PCR and Western blot were conducted to determine (A) the CCNA2 mRNA and (B) protein levels of CCNA2 as well as p53. MTT, Transwell, and flow cytometry were performed to investigate the effects of silencing CCNA2 on EC cell activities, including (C) cell viability, (D) migration and invasion, (E) cell cycle. * P < 0.05.

Article Snippet: In order to investigate the underlying mechanism of CCNA2 on EC cells, cells were classified into four groups: si-NC + DMSO, si-CCNA2 + DMSO, si-NC + PFTβand si-CCNA2 + PFTβ groups [PFTβ, p53 inhibitor, HY-16702, MedChemExpress, 10 μM ( )]. qRT-PCR was firstly performed to test CCNA2 level in each group, finding that CCNA2 was markedly decreased in cells transfected with si-CCNA2 + DMSO ( ).

Techniques: Migration, Transfection, Quantitative RT-PCR, Western Blot, Flow Cytometry

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: MiR-29c-3p Suppresses the Migration, Invasion and Cell Cycle in Esophageal Carcinoma via CCNA2/p53 Axis

doi: 10.3389/fbioe.2020.00075

Figure Lengend Snippet: MiR-29c-3p mediates the migration, invasion and cell cycle in EC via CCNA2/p53 axis. Cells were treated with inhibitor NC + si-NC, inhibitor NC + si-CCNA2, miR-29c-3p inhibitor + si-NC and miR-29c-3p inhibitor + si-CCNA2, and then harvested for (A) Western blot to detect the protein levels of CCNA2 and p53. (B) MTT was performed to test cell viability, (C) Transwell was conducted to assay the ability of cell migration and invasion, and (D) flow cytometry was carried out to determine the effect of miR-29c-3p on cell cycle. * P < 0.05.

Article Snippet: In order to investigate the underlying mechanism of CCNA2 on EC cells, cells were classified into four groups: si-NC + DMSO, si-CCNA2 + DMSO, si-NC + PFTβand si-CCNA2 + PFTβ groups [PFTβ, p53 inhibitor, HY-16702, MedChemExpress, 10 μM ( )]. qRT-PCR was firstly performed to test CCNA2 level in each group, finding that CCNA2 was markedly decreased in cells transfected with si-CCNA2 + DMSO ( ).

Techniques: Migration, Western Blot, Flow Cytometry

Journal: International immunopharmacology

Article Title: Immunosensitivity mediated by downregulated AKT1-SKP2 induces anti-PD-1-associated thyroid immune injury.

doi: 10.1016/j.intimp.2023.110452

Figure Lengend Snippet: Fig. 5. The downstream regulation of AKT1-SKP2 in NTHY cells.

Article Snippet: SKP2 inhibitor (SKP2i, MCE, 5uM) is solubilized by DMSO.

Techniques:

Journal: International immunopharmacology

Article Title: Immunosensitivity mediated by downregulated AKT1-SKP2 induces anti-PD-1-associated thyroid immune injury.

doi: 10.1016/j.intimp.2023.110452

Figure Lengend Snippet: Fig. 6. Interaction between SKP2 and PD-L1. (A) DAPI is in blue, SKP2 in red, and PD-L1 in green. In NTHY cells, SKP2 is mainly expressed in the nucleus and PD- L1 in non-nuclear area. (B) NTHY cells are co-transfected with pcDNA3.1-SKP2 plasmid and pcDNA3.1-PDL1 plasmid. The interaction between SKP2 and PD-L1 protein is proved by immunoprecipitation.

Article Snippet: SKP2 inhibitor (SKP2i, MCE, 5uM) is solubilized by DMSO.

Techniques: Transfection, Plasmid Preparation, Immunoprecipitation

Journal: International immunopharmacology

Article Title: Immunosensitivity mediated by downregulated AKT1-SKP2 induces anti-PD-1-associated thyroid immune injury.

doi: 10.1016/j.intimp.2023.110452

Figure Lengend Snippet: Fig. 8. Hypothesis: NIVO increases the immunosensitivity of NTHY cells by downregulating AKT1-SKP2.

Article Snippet: SKP2 inhibitor (SKP2i, MCE, 5uM) is solubilized by DMSO.

Techniques:

Journal: International immunopharmacology

Article Title: Immunosensitivity mediated by downregulated AKT1-SKP2 induces anti-PD-1-associated thyroid immune injury.

doi: 10.1016/j.intimp.2023.110452

Figure Lengend Snippet: Fig. 7. AKT1, SKP2, PD-L1 and IFN-γ in thyroid tissue. (A) The thyroid gland of mice is used for detection by immunohistochemistry. Fewer AKT1, SKP2, PD-L1 and more IFN-γ are observed in the NIVO group than in the CTRL group (10X). (B) Further, levels of various proteins in mouse thyroid are quantified by WB.(C) Human thyroid tissue is used for detection by WB. results revealed that PD-L1 is down-regulated and IFNG is up-regulated in the HT group compared to the non-HT group, however, there is no difference in AKT1 and SKP2.

Article Snippet: SKP2 inhibitor (SKP2i, MCE, 5uM) is solubilized by DMSO.

Techniques: Immunohistochemistry

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

Journal: Cancer Cell International

Article Title: The antiproliferative effect of C 2 -ceramide on lung cancer cells through apoptosis by inhibiting Akt and NFκB

doi: 10.1186/1475-2867-14-1

Figure Lengend Snippet: p-Akt and p-NFκB levels of C 2 -ceramide-treated H1299 lung cancer cells. Cells treated with different concentrations (0 to 50 μM) of C 2 -ceramide for 24 h. After treatment, the protein lysates were resolved by SDS-PAGE, transferred onto nitrocellulose membranes and probed with specific antibodies and detected signals using an enhanced chemiluminescence kit. (a) The changes of Akt and NFκB phosphorylation. (b) The changes of protein level of survivin, cyclin A2 and Bax. β-actin as an internal control.

Article Snippet: Subsequently, it was reacted with primary antibodies against t-Akt (#1081), p-NFκB (Ser536, #2220) and Bax (#1063, Epitomics, CA, USA); t-NFκB (sc-8008), β-catenin (sc-7963) and p-Akt (Ser473, sc-7985, Santa Cruz Biotech, Santa Cruz, CA, USA); Cyclin A2 (GeneTex Co., Cat No. GTX103042); survivin (AnaSpec, San Jose, CA, USA) and β-actin (#sc-8432, Santa Cruz Biotech), and their corresponding secondary antibodies.

Techniques: SDS Page, Phospho-proteomics, Control

Journal: Cancer Cell International

Article Title: The antiproliferative effect of C 2 -ceramide on lung cancer cells through apoptosis by inhibiting Akt and NFκB

doi: 10.1186/1475-2867-14-1

Figure Lengend Snippet: Schematic diagram of hypothesized mechanism of C 2 -ceramide-induced apoptosis of lung cancer cells. C 2 -ceramide inhibits the activity of both Akt and NF-κB, causing the down-regulation of pro-survival survivin and cell cycle promoter cyclin A2. On the contrary, C 2 -ceramide increases the protein level of pro-apoptotic Bax. As a result, C 2 -creamide treatment causes cell cycle G 1 arrest and chromatin condensation, subsequently, triggering the apoptosis of lung cancer cells.

Article Snippet: Subsequently, it was reacted with primary antibodies against t-Akt (#1081), p-NFκB (Ser536, #2220) and Bax (#1063, Epitomics, CA, USA); t-NFκB (sc-8008), β-catenin (sc-7963) and p-Akt (Ser473, sc-7985, Santa Cruz Biotech, Santa Cruz, CA, USA); Cyclin A2 (GeneTex Co., Cat No. GTX103042); survivin (AnaSpec, San Jose, CA, USA) and β-actin (#sc-8432, Santa Cruz Biotech), and their corresponding secondary antibodies.

Techniques: Activity Assay