Journal: Cancers

Article Title: Association of Proteasome Activity and Pool Heterogeneity with Markers Determining the Molecular Subtypes of Breast Cancer

doi: 10.3390/cancers17010159

Figure Lengend Snippet: Proteasome activities and content in BC and adjacent tissues. Representative immunohistochemical staining of tumor tissues ( a ). First row ( A1 – A4 ) Invasive carcinoma of nonspecific type (Luminal A subtype): A1 —Strong expression of ER; A2 —Strong expression of PR; A3 —negative expression of HER-2; A4 —low level of Ki-67. Second row ( B1 – B4 ) Invasive carcinoma of nonspecific type (Luminal B subtype): B1 —Strong expression of ER; B2 —Strong expression of PR; B3 —weak (1+) expression of HER-2; A4 —high level of Ki-67 (>20%). Third row ( C1 – C4 ) Invasive carcinoma of nonspecific type (Triple negative subtype): C1 —negative expression of ER; C2 , negative expression of PR; C3 , negative expression of HER-2; A4 , high level of Ki-67 (>50%). Immunohistochemistry, ×200. Scale bar—100 μm. ( b ) ChTL and CL proteasome activities in BC and adjacent tissues ( c ). Proteasome activities in tissues of various molecular subtypes of BC. ( d ) The content of 20S proteasome or proteasome regulator subunits ((α1, α2, α3, α5, α6, α7), Rpt6, PA28β LMP-2, LMP-7) determined by Western blotting in BC (1) and adjacent tissues (2). Representative Western blots are shown. ( e ) Content of proteasome subunits in tumour and adjacent tissue; and in different molecular subtypes of the BC ( f ) determined via quantification of blots using the ImageJ software ver. 1.54i. Red box indicates the relative content of proteasome subunits in the adjacent tissue, which was considered as 100%; * p < 0.05, ** p < 0.01, Mann-Whitney test.

Article Snippet: Primary mouse monoclonal antibodies to 20S proteasome subunits α1, α2, α3, α5, α6, α7 (cat. # sc-58412), mouse mAbs to LMP-7 (cat. # sc-100284), rabbit polyclonal antibodies (pAbs) to LMP-2 (cat. # sc-28809), and goat antibodies to the subunit PA28β (cat. # sc-23642) were obtained from Santa Cruz Biotechnology, Dallas, TX, USA.

Techniques: Immunohistochemical staining, Staining, Expressing, Immunohistochemistry, Western Blot, Software, MANN-WHITNEY

Journal: Cancers

Article Title: Association of Proteasome Activity and Pool Heterogeneity with Markers Determining the Molecular Subtypes of Breast Cancer

doi: 10.3390/cancers17010159

Figure Lengend Snippet: Relative content of proteasome subunits in tumor tissue in different molecular subtypes of BC.

Article Snippet: Primary mouse monoclonal antibodies to 20S proteasome subunits α1, α2, α3, α5, α6, α7 (cat. # sc-58412), mouse mAbs to LMP-7 (cat. # sc-100284), rabbit polyclonal antibodies (pAbs) to LMP-2 (cat. # sc-28809), and goat antibodies to the subunit PA28β (cat. # sc-23642) were obtained from Santa Cruz Biotechnology, Dallas, TX, USA.

Techniques:

Journal: PLoS ONE

Article Title: Immunoproteasome Overexpression Underlies the Pathogenesis of Thyroid Oncocytes and Primary Hypothyroidism: Studies in Humans and Mice

doi: 10.1371/journal.pone.0007857

Figure Lengend Snippet: Comparison of gene expression by SAGE in thyrocytes isolated from thyr -IFNγ transgenic (TG) or wild type (WT) mice (shows the genes that displayed the highest TG over WT ratio).

Article Snippet: After blocking aspecific binding with 5% normal goat serum, sections were incubated with the primary antibody; rabbit anti proteasome subunit (LMP2, LMP7, LMP10, PA28α, and PA28β; Biomol International, L.P., Plymouth Meeting, PA), in PBS supplemented with 1% bovine serum albumin, and incubated overnight at 4°C in a humid chamber.

Techniques: Comparison, Gene Expression, Isolation, Transgenic Assay, Membrane

Journal: PLoS ONE

Article Title: Hepatic Deficiency of COP9 Signalosome Subunit 8 Induces Ubiquitin-Proteasome System Impairment and Bim-Mediated Apoptosis in Murine Livers

doi: 10.1371/journal.pone.0067793

Figure Lengend Snippet: Four pairs of HS-Csn8KO and CTL littermate mice at 4-week-old were used for analyses presented here. ( A ) Representative images of western blot analyses of the indicated Bcl2 family proteins. β-Tubulin was probed as loading control (B~D). Summary of quantitative analyses. Pooled densitometry data of Bim ( B ), Bax ( C ), Bcl2 and Bcl-xL ( D ) normalized to β-tubulin are presented. *p<0.05, **p<0.01 vs. CTL; n=4 mice/group.

Article Snippet: Western blots were performed with following antibodies: Rpn2, Rpn8, Rpn10, Rpt5, PA28α, PA28β, PA28γ, VHL (von Hippel–Lindau), HIF1α (hypoxia inducible factor 1α), and Csn8 (Biomol), Bim and Bcl-xL (Chemicon), Bax and Bcl2 (BD Pharmingen Tech.), Nedd8 (Alexis), β-tubulin and ubiquitin (Sigma), CIS (cytokine- i nducible Src homology 2 domain-containing protein) and GFP (Santa Cruz Biotechnology), and 20S subunit β5 (custom made).

Techniques: Western Blot

Journal: PLoS ONE

Article Title: Hepatic Deficiency of COP9 Signalosome Subunit 8 Induces Ubiquitin-Proteasome System Impairment and Bim-Mediated Apoptosis in Murine Livers

doi: 10.1371/journal.pone.0067793

Figure Lengend Snippet: Crude proteins extracts from liver tissues were used for immunoprecipitation (IP) followed by iimunoblotting (IB) analyses. ( A ) Immunoprecipitation using a Bim-specific antibody followed by IB for the indicated proteins. ( B ) Pooled densitometry data of the IP western blot analyses as illustrated in panel A. For each protein, the density of the Csn8KO group is shown as the relative value to the average value of the corresponding CTL group. Arbitrary unit is used. ( C ) Immunoprecipitation of Bcl2 was followed by IB for Bcl2, Bim, and Bax. NS denotes a non-specific band. ( D ) Pooled densitometry data of the IP western blot analyses as illustrated in panel C. ( E ) Immunoprecipitation of Bax was followed by IB for Bax and Bim. *p<0.05, **p<0.01 vs. CTL.

Article Snippet: Western blots were performed with following antibodies: Rpn2, Rpn8, Rpn10, Rpt5, PA28α, PA28β, PA28γ, VHL (von Hippel–Lindau), HIF1α (hypoxia inducible factor 1α), and Csn8 (Biomol), Bim and Bcl-xL (Chemicon), Bax and Bcl2 (BD Pharmingen Tech.), Nedd8 (Alexis), β-tubulin and ubiquitin (Sigma), CIS (cytokine- i nducible Src homology 2 domain-containing protein) and GFP (Santa Cruz Biotechnology), and 20S subunit β5 (custom made).

Techniques: Immunoprecipitation, Western Blot

Journal: PLoS ONE

Article Title: Hepatic Deficiency of COP9 Signalosome Subunit 8 Induces Ubiquitin-Proteasome System Impairment and Bim-Mediated Apoptosis in Murine Livers

doi: 10.1371/journal.pone.0067793

Figure Lengend Snippet: Four pairs of HS-Csn8KO and CTL littermate mice at 4-week-old were used for analyses presented here. ( A ) Representative images of western blot analyses of the indicated Bcl2 family proteins. β-Tubulin was probed as loading control (B~D). Summary of quantitative analyses. Pooled densitometry data of Bim ( B ), Bax ( C ), Bcl2 and Bcl-xL ( D ) normalized to β-tubulin are presented. *p<0.05, **p<0.01 vs. CTL; n=4 mice/group.

Article Snippet: Western blots were performed with following antibodies: Rpn2, Rpn8, Rpn10, Rpt5, PA28α, PA28β, PA28γ, VHL (von Hippel–Lindau), HIF1α (hypoxia inducible factor 1α), and Csn8 (Biomol), Bim and Bcl-xL (Chemicon), Bax and Bcl2 (BD Pharmingen Tech.), Nedd8 (Alexis), β-tubulin and ubiquitin (Sigma), CIS (cytokine- i nducible Src homology 2 domain-containing protein) and GFP (Santa Cruz Biotechnology), and 20S subunit β5 (custom made).

Techniques: Western Blot

Journal: PLoS ONE

Article Title: Hepatic Deficiency of COP9 Signalosome Subunit 8 Induces Ubiquitin-Proteasome System Impairment and Bim-Mediated Apoptosis in Murine Livers

doi: 10.1371/journal.pone.0067793

Figure Lengend Snippet: Crude proteins extracts from liver tissues were used for immunoprecipitation (IP) followed by iimunoblotting (IB) analyses. ( A ) Immunoprecipitation using a Bim-specific antibody followed by IB for the indicated proteins. ( B ) Pooled densitometry data of the IP western blot analyses as illustrated in panel A. For each protein, the density of the Csn8KO group is shown as the relative value to the average value of the corresponding CTL group. Arbitrary unit is used. ( C ) Immunoprecipitation of Bcl2 was followed by IB for Bcl2, Bim, and Bax. NS denotes a non-specific band. ( D ) Pooled densitometry data of the IP western blot analyses as illustrated in panel C. ( E ) Immunoprecipitation of Bax was followed by IB for Bax and Bim. *p<0.05, **p<0.01 vs. CTL.

Article Snippet: Western blots were performed with following antibodies: Rpn2, Rpn8, Rpn10, Rpt5, PA28α, PA28β, PA28γ, VHL (von Hippel–Lindau), HIF1α (hypoxia inducible factor 1α), and Csn8 (Biomol), Bim and Bcl-xL (Chemicon), Bax and Bcl2 (BD Pharmingen Tech.), Nedd8 (Alexis), β-tubulin and ubiquitin (Sigma), CIS (cytokine- i nducible Src homology 2 domain-containing protein) and GFP (Santa Cruz Biotechnology), and 20S subunit β5 (custom made).

Techniques: Immunoprecipitation, Western Blot