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: 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