Journal: Journal of Biological Chemistry

Article Title: IKKα Regulates Estrogen-induced Cell Cycle Progression by Modulating E2F1 Expression

doi: 10.1074/jbc.m512439200

Figure Lengend Snippet: FIGURE 2. IKK is required for the transcription of E2F1-responsive genes. MCF7 cells were transfected with control siRNA (40 nM) and siRNAs targeting either IKK (A) or IKK (B). At 24 h post- transfection, serum-free medium was added to the cells, and after 48 h of serum starvation, the cells were either untreated or treated with 100 nM E2fortheindicatedtimes.TotalRNAwasextracted from the cells, and the expression of IKK, IKK, E2F1, TK1, PCNA, cdc25A, cyclin E, and SKP2 was analyzed by real-time PCR and normalized to that of 18 S RNA. Representative results from three independent experiments are shown. Asterisks indicate statistical significance (p 0.01).

Article Snippet: The primers and probes included E2F1 (Hs00153451), IKK (Hs00175141), IKK (Hs00395088), TK1 (Hs00177406), PCNA (Hs00427214), cdc25A (Hs00153168), cyclin E (Hs00233356), cyclin D1 (Hs00277939), SKP2 (Hs00180634), and 18 S rRNA (4333760) as a reference gene and analyzed in real-time quantitative PCR obtained from Assays-on-Demand products (Applied Biosystems). siRNA Transfection—siRNAs targeting IKK and the techniques for transfection were described previously (46).

Techniques: Transfection, Control, Expressing, Real-time Polymerase Chain Reaction

Journal: Science signaling

Article Title: Skp2-dependent reactivation of AKT drives resistance to PI3K inhibitors

doi: 10.1126/scisignal.aao3810

Figure Lengend Snippet: (A) Representative images of MDA-MB-231 spheroids, from parental or BKM-120–resistant cells, that were infected with SKP2 shRNA lentiviral vector or control pLKO, and grown for 14 days in a 3-dimensional Matrigel/growth media mixture and treated with BKM-120 and/or MK-2206 (each 1 μM). Scale bar, 500 μm (4x magnification).

Article Snippet: After mixing the resulting template with Skp2 (Mm00449925_m1) or glyceraldehyde-3-phospahte dehydrogenase (GAPDH; Mm99999915_g1) primers and ABI Taqman Fast Universal PCR Master Mix, the RT-PCR reaction was performed with the ABI-7500 Fast Real-time PCR system.

Techniques: Infection, shRNA, Plasmid Preparation, Control

Journal: Science signaling

Article Title: Skp2-dependent reactivation of AKT drives resistance to PI3K inhibitors

doi: 10.1126/scisignal.aao3810

Figure Lengend Snippet: (A) Immunoblotting of lysates from MDA-MB-231, parental and BKM-120–resistant, cells that were infected with SKP2 shRNA lentiviral vector or control pLKO. Cells were serum-starved overnight (−) or stimulated with IGF-1 for 20 min (+).

Article Snippet: After mixing the resulting template with Skp2 (Mm00449925_m1) or glyceraldehyde-3-phospahte dehydrogenase (GAPDH; Mm99999915_g1) primers and ABI Taqman Fast Universal PCR Master Mix, the RT-PCR reaction was performed with the ABI-7500 Fast Real-time PCR system.

Techniques: Western Blot, Infection, shRNA, Plasmid Preparation, Control

Journal: British Journal of Cancer

Article Title: Gene expression profile and response to trastuzumab–docetaxel-based treatment in breast carcinoma

doi: 10.1038/sj.bjc.6605310

Figure Lengend Snippet: References and nucleotide sequences of primers and probes used in this study

Article Snippet: , SKP2 , NM_032637 , Hs01021867_m1.

Techniques:

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: Scientific Reports

Article Title: The heavy chain of 4F2 antigen promote prostate cancer progression via SKP-2

doi: 10.1038/s41598-021-90748-9

Figure Lengend Snippet: 4F2hc RNA-sequencing analysis. RNA-seq analysis was performed with si4F2hc (si4F2hc-1 and si4F2hc-2) and compared with a control group ( A ). The most down-regulated 1403 genes are selected, and Matascape is used for analysis of significantly related with GO terms ( B ). Si4F2hc concentration-dependent effect on candidate genes was assessed by real-time PCR ( C ). SiSKP2 (siSKP2-1 and siSKP2-2) inhibits C4-2 cell proliferation ( D ). Data represent three independent experiments with similar results. P-values were calculated by the Mann–Whitney U-test. *** p < 0.001.

Article Snippet: In this study, siRNAs 4F2 cell-surface antigen heavy chain (si4F2hc) (Stealth siRNAs: HSS109825 and HSS109826), siRNAs S-Phase Kinase Associated Protein 2 (siSKP2) (Stealth siRNAs: HSS185711 and HSS109780), siRNA Negative Control (Stealth RNAi, Thermo Fisher Scientific, MA, USA), Lipofectamine 3000 Transfection Reagent (Invitrogen), were used.

Techniques: RNA Sequencing Assay, Concentration Assay, Real-time Polymerase Chain Reaction, MANN-WHITNEY