anti α tubulin Search Results


sheep polyclonal anti tubulin  (Cytoskeleton Inc)
95
Cytoskeleton Inc sheep polyclonal anti tubulin
Sheep Polyclonal Anti Tubulin, supplied by Cytoskeleton Inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/anti+%CE%B1+tubulin/Anti-alpha+beta+tubulin+sheep+polyclonal/pmc12562719-60-40-44
Average 95 stars, based on 1 article reviews
sheep polyclonal anti tubulin - by Bioz Stars, 2026-10
95/100 stars
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tuj  (AvesLabs)
98
AvesLabs tuj
Tuj, supplied by AvesLabs, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/anti+%CE%B1+tubulin/Anti-Beta-Tubulin+3+(TUJ)+Antibody/pm39814010-263-82-78
Average 98 stars, based on 1 article reviews
tuj - by Bioz Stars, 2026-10
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tubulin  (Developmental Studies Hybridoma Bank)
98
Developmental Studies Hybridoma Bank tubulin
Spag protein structure and patterns of gene expression and protein accumulation during Drosophila development. A, schematic structure of the Spag protein and evolutionary conservation of the TPR region in Spag orthologs from yeast to human. Filled boxes represent predicted structural domains and lines unstructured domains. The three TPR motifs forming the TPR region are indicated as red, blue, and green boxes. TPR 3′ is a putative helical domain absent in Tah1. Orange box represents a potential α-helical junction between TPR 3 and TPR 3′. The potential monad binding motif (interPro 025986) overlaps with a predicted structured domain (magenta). Below are aligned the amino acid residues characterizing the three TPR motifs in S. cerevisiae (S.c.), D. melanogaster (D.m.), Danio rero (D.r.), Homo sapiens (H.s.), and Xenopus tropicalis (X.t.). Conserved residues identified in Tah1 for Hsp90 binding are indicated in bold red (21). Blue indicates additional residues conserved in TPR domains and important for Hsp70/Hsp90 binding, as defined previously (23). B, expression pattern of spag mRNA (left panel) and pattern of Spag protein accumulation (right panel) during Drosophila development. RNAs and proteins were extracted from (E) embryos with age in hours after egg laying indicated above the lanes (L1, L2, and L3) first, second, and third instar larvae, (P) pupae, (F) female, and (M) male 3-days old imagos. The Northern blot was hybridized with either 32P-labeled spag or <t>β-tubulin</t> cDNA. The Western blot was probed with anti-Spag or anti-ribosomal P40 <t>protein</t> <t>polyclonal</t> antibodies. The β-tubulin cDNA and anti-P40 antibodies were used for loading control.
Tubulin, supplied by Developmental Studies Hybridoma Bank, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/anti+%CE%B1+tubulin/anti-tubulin/pmc03937688-112-65-69
Average 98 stars, based on 1 article reviews
tubulin - by Bioz Stars, 2026-10
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hfabtm rhodamine anti tubulin  (Bio-Rad)
96
Bio-Rad hfabtm rhodamine anti tubulin
Spag protein structure and patterns of gene expression and protein accumulation during Drosophila development. A, schematic structure of the Spag protein and evolutionary conservation of the TPR region in Spag orthologs from yeast to human. Filled boxes represent predicted structural domains and lines unstructured domains. The three TPR motifs forming the TPR region are indicated as red, blue, and green boxes. TPR 3′ is a putative helical domain absent in Tah1. Orange box represents a potential α-helical junction between TPR 3 and TPR 3′. The potential monad binding motif (interPro 025986) overlaps with a predicted structured domain (magenta). Below are aligned the amino acid residues characterizing the three TPR motifs in S. cerevisiae (S.c.), D. melanogaster (D.m.), Danio rero (D.r.), Homo sapiens (H.s.), and Xenopus tropicalis (X.t.). Conserved residues identified in Tah1 for Hsp90 binding are indicated in bold red (21). Blue indicates additional residues conserved in TPR domains and important for Hsp70/Hsp90 binding, as defined previously (23). B, expression pattern of spag mRNA (left panel) and pattern of Spag protein accumulation (right panel) during Drosophila development. RNAs and proteins were extracted from (E) embryos with age in hours after egg laying indicated above the lanes (L1, L2, and L3) first, second, and third instar larvae, (P) pupae, (F) female, and (M) male 3-days old imagos. The Northern blot was hybridized with either 32P-labeled spag or <t>β-tubulin</t> cDNA. The Western blot was probed with anti-Spag or anti-ribosomal P40 <t>protein</t> <t>polyclonal</t> antibodies. The β-tubulin cDNA and anti-P40 antibodies were used for loading control.
Hfabtm Rhodamine Anti Tubulin, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/anti+%CE%B1+tubulin/hFAB+Rhodamine+Anti-Tubulin+Primary+Antibody/bio_rxiv__2025__08__04__668513-294-42-46
Average 96 stars, based on 1 article reviews
hfabtm rhodamine anti tubulin - by Bioz Stars, 2026-10
96/100 stars
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equal loading  (Bio-Rad)
94
Bio-Rad equal loading
Spag protein structure and patterns of gene expression and protein accumulation during Drosophila development. A, schematic structure of the Spag protein and evolutionary conservation of the TPR region in Spag orthologs from yeast to human. Filled boxes represent predicted structural domains and lines unstructured domains. The three TPR motifs forming the TPR region are indicated as red, blue, and green boxes. TPR 3′ is a putative helical domain absent in Tah1. Orange box represents a potential α-helical junction between TPR 3 and TPR 3′. The potential monad binding motif (interPro 025986) overlaps with a predicted structured domain (magenta). Below are aligned the amino acid residues characterizing the three TPR motifs in S. cerevisiae (S.c.), D. melanogaster (D.m.), Danio rero (D.r.), Homo sapiens (H.s.), and Xenopus tropicalis (X.t.). Conserved residues identified in Tah1 for Hsp90 binding are indicated in bold red (21). Blue indicates additional residues conserved in TPR domains and important for Hsp70/Hsp90 binding, as defined previously (23). B, expression pattern of spag mRNA (left panel) and pattern of Spag protein accumulation (right panel) during Drosophila development. RNAs and proteins were extracted from (E) embryos with age in hours after egg laying indicated above the lanes (L1, L2, and L3) first, second, and third instar larvae, (P) pupae, (F) female, and (M) male 3-days old imagos. The Northern blot was hybridized with either 32P-labeled spag or <t>β-tubulin</t> cDNA. The Western blot was probed with anti-Spag or anti-ribosomal P40 <t>protein</t> <t>polyclonal</t> antibodies. The β-tubulin cDNA and anti-P40 antibodies were used for loading control.
Equal Loading, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/anti+%CE%B1+tubulin/Rabbit+anti+Tubulin/pmc04795661-56-11-26
Average 94 stars, based on 1 article reviews
equal loading - by Bioz Stars, 2026-10
94/100 stars
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rat anti tubulin primary antibody  (Bio-Rad)
96
Bio-Rad rat anti tubulin primary antibody
Spag protein structure and patterns of gene expression and protein accumulation during Drosophila development. A, schematic structure of the Spag protein and evolutionary conservation of the TPR region in Spag orthologs from yeast to human. Filled boxes represent predicted structural domains and lines unstructured domains. The three TPR motifs forming the TPR region are indicated as red, blue, and green boxes. TPR 3′ is a putative helical domain absent in Tah1. Orange box represents a potential α-helical junction between TPR 3 and TPR 3′. The potential monad binding motif (interPro 025986) overlaps with a predicted structured domain (magenta). Below are aligned the amino acid residues characterizing the three TPR motifs in S. cerevisiae (S.c.), D. melanogaster (D.m.), Danio rero (D.r.), Homo sapiens (H.s.), and Xenopus tropicalis (X.t.). Conserved residues identified in Tah1 for Hsp90 binding are indicated in bold red (21). Blue indicates additional residues conserved in TPR domains and important for Hsp70/Hsp90 binding, as defined previously (23). B, expression pattern of spag mRNA (left panel) and pattern of Spag protein accumulation (right panel) during Drosophila development. RNAs and proteins were extracted from (E) embryos with age in hours after egg laying indicated above the lanes (L1, L2, and L3) first, second, and third instar larvae, (P) pupae, (F) female, and (M) male 3-days old imagos. The Northern blot was hybridized with either 32P-labeled spag or <t>β-tubulin</t> cDNA. The Western blot was probed with anti-Spag or anti-ribosomal P40 <t>protein</t> <t>polyclonal</t> antibodies. The β-tubulin cDNA and anti-P40 antibodies were used for loading control.
Rat Anti Tubulin Primary Antibody, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/anti+%CE%B1+tubulin/Rat+anti+Tubulin+Alpha/pmc13065327-344-1-5
Average 96 stars, based on 1 article reviews
rat anti tubulin primary antibody - by Bioz Stars, 2026-10
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α tubulin  (Cedarlane)
93
Cedarlane α tubulin
Spag protein structure and patterns of gene expression and protein accumulation during Drosophila development. A, schematic structure of the Spag protein and evolutionary conservation of the TPR region in Spag orthologs from yeast to human. Filled boxes represent predicted structural domains and lines unstructured domains. The three TPR motifs forming the TPR region are indicated as red, blue, and green boxes. TPR 3′ is a putative helical domain absent in Tah1. Orange box represents a potential α-helical junction between TPR 3 and TPR 3′. The potential monad binding motif (interPro 025986) overlaps with a predicted structured domain (magenta). Below are aligned the amino acid residues characterizing the three TPR motifs in S. cerevisiae (S.c.), D. melanogaster (D.m.), Danio rero (D.r.), Homo sapiens (H.s.), and Xenopus tropicalis (X.t.). Conserved residues identified in Tah1 for Hsp90 binding are indicated in bold red (21). Blue indicates additional residues conserved in TPR domains and important for Hsp70/Hsp90 binding, as defined previously (23). B, expression pattern of spag mRNA (left panel) and pattern of Spag protein accumulation (right panel) during Drosophila development. RNAs and proteins were extracted from (E) embryos with age in hours after egg laying indicated above the lanes (L1, L2, and L3) first, second, and third instar larvae, (P) pupae, (F) female, and (M) male 3-days old imagos. The Northern blot was hybridized with either 32P-labeled spag or <t>β-tubulin</t> cDNA. The Western blot was probed with anti-Spag or anti-ribosomal P40 <t>protein</t> <t>polyclonal</t> antibodies. The β-tubulin cDNA and anti-P40 antibodies were used for loading control.
α Tubulin, supplied by Cedarlane, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/anti+%CE%B1+tubulin/Anti-Human+alpha-Tubulin%2C+Ascites%2C+(Clone+DM1A)+(mouse+IgG1)/pm40299057-92-30-32
Average 93 stars, based on 1 article reviews
α tubulin - by Bioz Stars, 2026-10
93/100 stars
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primary antibody polyclonal rabbit anti α tubulin antiserum  (Rockland Immunochemicals)
93
Rockland Immunochemicals primary antibody polyclonal rabbit anti α tubulin antiserum
Spag protein structure and patterns of gene expression and protein accumulation during Drosophila development. A, schematic structure of the Spag protein and evolutionary conservation of the TPR region in Spag orthologs from yeast to human. Filled boxes represent predicted structural domains and lines unstructured domains. The three TPR motifs forming the TPR region are indicated as red, blue, and green boxes. TPR 3′ is a putative helical domain absent in Tah1. Orange box represents a potential α-helical junction between TPR 3 and TPR 3′. The potential monad binding motif (interPro 025986) overlaps with a predicted structured domain (magenta). Below are aligned the amino acid residues characterizing the three TPR motifs in S. cerevisiae (S.c.), D. melanogaster (D.m.), Danio rero (D.r.), Homo sapiens (H.s.), and Xenopus tropicalis (X.t.). Conserved residues identified in Tah1 for Hsp90 binding are indicated in bold red (21). Blue indicates additional residues conserved in TPR domains and important for Hsp70/Hsp90 binding, as defined previously (23). B, expression pattern of spag mRNA (left panel) and pattern of Spag protein accumulation (right panel) during Drosophila development. RNAs and proteins were extracted from (E) embryos with age in hours after egg laying indicated above the lanes (L1, L2, and L3) first, second, and third instar larvae, (P) pupae, (F) female, and (M) male 3-days old imagos. The Northern blot was hybridized with either 32P-labeled spag or <t>β-tubulin</t> cDNA. The Western blot was probed with anti-Spag or anti-ribosomal P40 <t>protein</t> <t>polyclonal</t> antibodies. The β-tubulin cDNA and anti-P40 antibodies were used for loading control.
Primary Antibody Polyclonal Rabbit Anti α Tubulin Antiserum, supplied by Rockland Immunochemicals, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/anti+%CE%B1+tubulin/AFFINITY+PURF%2E+ANTI-ALPHA-TUBULIN(RABBIT/pmc07845644-300-6-15
Average 93 stars, based on 1 article reviews
primary antibody polyclonal rabbit anti α tubulin antiserum - by Bioz Stars, 2026-10
93/100 stars
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gephyrin  (Alomone Labs)
94
Alomone Labs gephyrin
Spag protein structure and patterns of gene expression and protein accumulation during Drosophila development. A, schematic structure of the Spag protein and evolutionary conservation of the TPR region in Spag orthologs from yeast to human. Filled boxes represent predicted structural domains and lines unstructured domains. The three TPR motifs forming the TPR region are indicated as red, blue, and green boxes. TPR 3′ is a putative helical domain absent in Tah1. Orange box represents a potential α-helical junction between TPR 3 and TPR 3′. The potential monad binding motif (interPro 025986) overlaps with a predicted structured domain (magenta). Below are aligned the amino acid residues characterizing the three TPR motifs in S. cerevisiae (S.c.), D. melanogaster (D.m.), Danio rero (D.r.), Homo sapiens (H.s.), and Xenopus tropicalis (X.t.). Conserved residues identified in Tah1 for Hsp90 binding are indicated in bold red (21). Blue indicates additional residues conserved in TPR domains and important for Hsp70/Hsp90 binding, as defined previously (23). B, expression pattern of spag mRNA (left panel) and pattern of Spag protein accumulation (right panel) during Drosophila development. RNAs and proteins were extracted from (E) embryos with age in hours after egg laying indicated above the lanes (L1, L2, and L3) first, second, and third instar larvae, (P) pupae, (F) female, and (M) male 3-days old imagos. The Northern blot was hybridized with either 32P-labeled spag or <t>β-tubulin</t> cDNA. The Western blot was probed with anti-Spag or anti-ribosomal P40 <t>protein</t> <t>polyclonal</t> antibodies. The β-tubulin cDNA and anti-P40 antibodies were used for loading control.
Gephyrin, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/anti+%CE%B1+tubulin/Anti-Gephyrin+Antibody/pmc12392074-22-0-8
Average 94 stars, based on 1 article reviews
gephyrin - by Bioz Stars, 2026-10
94/100 stars
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ferredoxin 1 fdx1  (Boster Bio)
93
Boster Bio ferredoxin 1 fdx1
Spag protein structure and patterns of gene expression and protein accumulation during Drosophila development. A, schematic structure of the Spag protein and evolutionary conservation of the TPR region in Spag orthologs from yeast to human. Filled boxes represent predicted structural domains and lines unstructured domains. The three TPR motifs forming the TPR region are indicated as red, blue, and green boxes. TPR 3′ is a putative helical domain absent in Tah1. Orange box represents a potential α-helical junction between TPR 3 and TPR 3′. The potential monad binding motif (interPro 025986) overlaps with a predicted structured domain (magenta). Below are aligned the amino acid residues characterizing the three TPR motifs in S. cerevisiae (S.c.), D. melanogaster (D.m.), Danio rero (D.r.), Homo sapiens (H.s.), and Xenopus tropicalis (X.t.). Conserved residues identified in Tah1 for Hsp90 binding are indicated in bold red (21). Blue indicates additional residues conserved in TPR domains and important for Hsp70/Hsp90 binding, as defined previously (23). B, expression pattern of spag mRNA (left panel) and pattern of Spag protein accumulation (right panel) during Drosophila development. RNAs and proteins were extracted from (E) embryos with age in hours after egg laying indicated above the lanes (L1, L2, and L3) first, second, and third instar larvae, (P) pupae, (F) female, and (M) male 3-days old imagos. The Northern blot was hybridized with either 32P-labeled spag or <t>β-tubulin</t> cDNA. The Western blot was probed with anti-Spag or anti-ribosomal P40 <t>protein</t> <t>polyclonal</t> antibodies. The β-tubulin cDNA and anti-P40 antibodies were used for loading control.
Ferredoxin 1 Fdx1, supplied by Boster Bio, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/anti+%CE%B1+tubulin/Anti-ADX+Rabbit+Monoclonal+Antibody/10__1016_slash_j__jrras__2025__102076-107-23-27
Average 93 stars, based on 1 article reviews
ferredoxin 1 fdx1 - by Bioz Stars, 2026-10
93/100 stars
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monoclonal mouse β tubulin iii  (Bio-Rad)
94
Bio-Rad monoclonal mouse β tubulin iii
Spag protein structure and patterns of gene expression and protein accumulation during Drosophila development. A, schematic structure of the Spag protein and evolutionary conservation of the TPR region in Spag orthologs from yeast to human. Filled boxes represent predicted structural domains and lines unstructured domains. The three TPR motifs forming the TPR region are indicated as red, blue, and green boxes. TPR 3′ is a putative helical domain absent in Tah1. Orange box represents a potential α-helical junction between TPR 3 and TPR 3′. The potential monad binding motif (interPro 025986) overlaps with a predicted structured domain (magenta). Below are aligned the amino acid residues characterizing the three TPR motifs in S. cerevisiae (S.c.), D. melanogaster (D.m.), Danio rero (D.r.), Homo sapiens (H.s.), and Xenopus tropicalis (X.t.). Conserved residues identified in Tah1 for Hsp90 binding are indicated in bold red (21). Blue indicates additional residues conserved in TPR domains and important for Hsp70/Hsp90 binding, as defined previously (23). B, expression pattern of spag mRNA (left panel) and pattern of Spag protein accumulation (right panel) during Drosophila development. RNAs and proteins were extracted from (E) embryos with age in hours after egg laying indicated above the lanes (L1, L2, and L3) first, second, and third instar larvae, (P) pupae, (F) female, and (M) male 3-days old imagos. The Northern blot was hybridized with either 32P-labeled spag or <t>β-tubulin</t> cDNA. The Western blot was probed with anti-Spag or anti-ribosomal P40 <t>protein</t> <t>polyclonal</t> antibodies. The β-tubulin cDNA and anti-P40 antibodies were used for loading control.
Monoclonal Mouse β Tubulin Iii, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/anti+%CE%B1+tubulin/Mouse+anti+Tubulin+Beta+3/pmc06622038-300-12-18
Average 94 stars, based on 1 article reviews
monoclonal mouse β tubulin iii - by Bioz Stars, 2026-10
94/100 stars
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tubulin  (Boster Bio)
90
Boster Bio tubulin
The SCN5A p.D372H variant <t>reduces</t> <t>Nav1.5</t> expression through decreased mRNA abundance and stability, independent of plasmid DNA levels. ( A ) Relative SCN5A mRNA expression measured by RT-qPCR. Transcript levels were significantly reduced in the D372H group compared with WT (**** p < 0.0001). Co-expression of WT and D372H (WT+MUT) also resulted in a significant decrease in SCN5A mRNA expression compared with WT (**** p < 0.0001). Data were normalized to WT. ( B ) Quantification of intracellular SCN5A plasmid DNA levels. No significant differences were observed among WT, WT+MUT, and MUT groups (ns), indicating comparable transfection efficiency. ( C ) SCN5A mRNA decay following transcriptional inhibition with actinomycin D. Relative mRNA remaining was normalized to the 0-h time point. The D372H group exhibited accelerated mRNA decay compared with WT, indicating reduced transcript stability. ( D ) Representative Western blot images showing total Nav1.5 protein expression in BLANK, CONTROL, WT, WT+MUT, and MUT groups. Molecular weight markers are indicated in kilodaltons (kDa). The predicted molecular weight of native Nav1.5 is approximately 227 kDa; however, in this study SCN5A was expressed as a GFP-tagged fusion protein, resulting in an apparent band migrating above 245 kDa. <t>Tubulin</t> (50 kDa) was used as a loading control. ( E ) Quantitative analysis of Nav1.5 protein expression normalized to Tubulin. Protein levels were significantly reduced in the D372H group compared with WT (* p < 0.05). The WT+MUT group also showed decreased expression compared with WT (** p < 0.01). Data are presented as mean ± SD from at least three independent experiments.
Tubulin, supplied by Boster Bio, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/anti+%CE%B1+tubulin/Anti-Tubulin+alpha+Antibody+Picoband/pmc13024415-106-17-21
Average 90 stars, based on 1 article reviews
tubulin - by Bioz Stars, 2026-10
90/100 stars
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Image Search Results


Spag protein structure and patterns of gene expression and protein accumulation during Drosophila development. A, schematic structure of the Spag protein and evolutionary conservation of the TPR region in Spag orthologs from yeast to human. Filled boxes represent predicted structural domains and lines unstructured domains. The three TPR motifs forming the TPR region are indicated as red, blue, and green boxes. TPR 3′ is a putative helical domain absent in Tah1. Orange box represents a potential α-helical junction between TPR 3 and TPR 3′. The potential monad binding motif (interPro 025986) overlaps with a predicted structured domain (magenta). Below are aligned the amino acid residues characterizing the three TPR motifs in S. cerevisiae (S.c.), D. melanogaster (D.m.), Danio rero (D.r.), Homo sapiens (H.s.), and Xenopus tropicalis (X.t.). Conserved residues identified in Tah1 for Hsp90 binding are indicated in bold red (21). Blue indicates additional residues conserved in TPR domains and important for Hsp70/Hsp90 binding, as defined previously (23). B, expression pattern of spag mRNA (left panel) and pattern of Spag protein accumulation (right panel) during Drosophila development. RNAs and proteins were extracted from (E) embryos with age in hours after egg laying indicated above the lanes (L1, L2, and L3) first, second, and third instar larvae, (P) pupae, (F) female, and (M) male 3-days old imagos. The Northern blot was hybridized with either 32P-labeled spag or β-tubulin cDNA. The Western blot was probed with anti-Spag or anti-ribosomal P40 protein polyclonal antibodies. The β-tubulin cDNA and anti-P40 antibodies were used for loading control.

Journal: The Journal of Biological Chemistry

Article Title: Drosophila Spag Is the Homolog of RNA Polymerase II-associated Protein 3 (RPAP3) and Recruits the Heat Shock Proteins 70 and 90 (Hsp70 and Hsp90) during the Assembly of Cellular Machineries * ♦

doi: 10.1074/jbc.M113.499608

Figure Lengend Snippet: Spag protein structure and patterns of gene expression and protein accumulation during Drosophila development. A, schematic structure of the Spag protein and evolutionary conservation of the TPR region in Spag orthologs from yeast to human. Filled boxes represent predicted structural domains and lines unstructured domains. The three TPR motifs forming the TPR region are indicated as red, blue, and green boxes. TPR 3′ is a putative helical domain absent in Tah1. Orange box represents a potential α-helical junction between TPR 3 and TPR 3′. The potential monad binding motif (interPro 025986) overlaps with a predicted structured domain (magenta). Below are aligned the amino acid residues characterizing the three TPR motifs in S. cerevisiae (S.c.), D. melanogaster (D.m.), Danio rero (D.r.), Homo sapiens (H.s.), and Xenopus tropicalis (X.t.). Conserved residues identified in Tah1 for Hsp90 binding are indicated in bold red (21). Blue indicates additional residues conserved in TPR domains and important for Hsp70/Hsp90 binding, as defined previously (23). B, expression pattern of spag mRNA (left panel) and pattern of Spag protein accumulation (right panel) during Drosophila development. RNAs and proteins were extracted from (E) embryos with age in hours after egg laying indicated above the lanes (L1, L2, and L3) first, second, and third instar larvae, (P) pupae, (F) female, and (M) male 3-days old imagos. The Northern blot was hybridized with either 32P-labeled spag or β-tubulin cDNA. The Western blot was probed with anti-Spag or anti-ribosomal P40 protein polyclonal antibodies. The β-tubulin cDNA and anti-P40 antibodies were used for loading control.

Article Snippet: Proteins were detected as follows: Rpb1 detected with mouse monoclonal PB7-C2 antibody; Rpb2 with goat S20 from Santa Cruz Biotechnology; Nop58 with polyclonal antibodies generated from rabbits immunized with an KKLQEVDSLWKEFETPEK peptide ( 14 ); p70 S6K with monoclonal antibody SC-9027 from Santa Cruz Biotechnology; phospho-Thr-398 p70 S6K with monoclonal antibody provided by Cell Signaling Technology (reference 9209); fibrillarin with monoclonal antibody 5821 from Abcam; tubulin with monoclonal 12G10 (Developmental Studies Hybridoma Bank, Iowa City, IA); polyclonal anti-15.5K antibodies from Santa Cruz Biotechnology (SC-86760); polyclonal anti-P40 antibodies have been described previously ( 15 ), and anti-FLAG M2 monoclonal antibody was from Sigma.

Techniques: Gene Expression, Binding Assay, Expressing, Northern Blot, Labeling, Western Blot, Control

Spag and Nufip are required for box C/D sno core protein stabilization. A, Western blot analysis of protein extracts from third instar larvae spagk12101, as compared with wild-type w1118 (Ct), showed a significant diminution in the content of Nop58 (Nop5) and 15.5K (Hoip) but not fibrillarin. Tubulin was used as a loading control. B, this phenomenon was also observed in pupae extracts from animals in which RNAi was induced by Gal4act5C to knock down spag (Gal4act5c/spagRNAi as compared with Gal4act5c/+). C, similar results were observed in pupae extracts from animals in which RNAi was induced against Nufip (C, Gal4act5c/nufipRNAi compared with w1118).

Journal: The Journal of Biological Chemistry

Article Title: Drosophila Spag Is the Homolog of RNA Polymerase II-associated Protein 3 (RPAP3) and Recruits the Heat Shock Proteins 70 and 90 (Hsp70 and Hsp90) during the Assembly of Cellular Machineries * ♦

doi: 10.1074/jbc.M113.499608

Figure Lengend Snippet: Spag and Nufip are required for box C/D sno core protein stabilization. A, Western blot analysis of protein extracts from third instar larvae spagk12101, as compared with wild-type w1118 (Ct), showed a significant diminution in the content of Nop58 (Nop5) and 15.5K (Hoip) but not fibrillarin. Tubulin was used as a loading control. B, this phenomenon was also observed in pupae extracts from animals in which RNAi was induced by Gal4act5C to knock down spag (Gal4act5c/spagRNAi as compared with Gal4act5c/+). C, similar results were observed in pupae extracts from animals in which RNAi was induced against Nufip (C, Gal4act5c/nufipRNAi compared with w1118).

Article Snippet: Proteins were detected as follows: Rpb1 detected with mouse monoclonal PB7-C2 antibody; Rpb2 with goat S20 from Santa Cruz Biotechnology; Nop58 with polyclonal antibodies generated from rabbits immunized with an KKLQEVDSLWKEFETPEK peptide ( 14 ); p70 S6K with monoclonal antibody SC-9027 from Santa Cruz Biotechnology; phospho-Thr-398 p70 S6K with monoclonal antibody provided by Cell Signaling Technology (reference 9209); fibrillarin with monoclonal antibody 5821 from Abcam; tubulin with monoclonal 12G10 (Developmental Studies Hybridoma Bank, Iowa City, IA); polyclonal anti-15.5K antibodies from Santa Cruz Biotechnology (SC-86760); polyclonal anti-P40 antibodies have been described previously ( 15 ), and anti-FLAG M2 monoclonal antibody was from Sigma.

Techniques: Western Blot, Control, Knockdown

The SCN5A p.D372H variant reduces Nav1.5 expression through decreased mRNA abundance and stability, independent of plasmid DNA levels. ( A ) Relative SCN5A mRNA expression measured by RT-qPCR. Transcript levels were significantly reduced in the D372H group compared with WT (**** p < 0.0001). Co-expression of WT and D372H (WT+MUT) also resulted in a significant decrease in SCN5A mRNA expression compared with WT (**** p < 0.0001). Data were normalized to WT. ( B ) Quantification of intracellular SCN5A plasmid DNA levels. No significant differences were observed among WT, WT+MUT, and MUT groups (ns), indicating comparable transfection efficiency. ( C ) SCN5A mRNA decay following transcriptional inhibition with actinomycin D. Relative mRNA remaining was normalized to the 0-h time point. The D372H group exhibited accelerated mRNA decay compared with WT, indicating reduced transcript stability. ( D ) Representative Western blot images showing total Nav1.5 protein expression in BLANK, CONTROL, WT, WT+MUT, and MUT groups. Molecular weight markers are indicated in kilodaltons (kDa). The predicted molecular weight of native Nav1.5 is approximately 227 kDa; however, in this study SCN5A was expressed as a GFP-tagged fusion protein, resulting in an apparent band migrating above 245 kDa. Tubulin (50 kDa) was used as a loading control. ( E ) Quantitative analysis of Nav1.5 protein expression normalized to Tubulin. Protein levels were significantly reduced in the D372H group compared with WT (* p < 0.05). The WT+MUT group also showed decreased expression compared with WT (** p < 0.01). Data are presented as mean ± SD from at least three independent experiments.

Journal: Biomedicines

Article Title: Functional Characterization of the SCN5A p.D372H Variant Associated with Brugada Syndrome

doi: 10.3390/biomedicines14030582

Figure Lengend Snippet: The SCN5A p.D372H variant reduces Nav1.5 expression through decreased mRNA abundance and stability, independent of plasmid DNA levels. ( A ) Relative SCN5A mRNA expression measured by RT-qPCR. Transcript levels were significantly reduced in the D372H group compared with WT (**** p < 0.0001). Co-expression of WT and D372H (WT+MUT) also resulted in a significant decrease in SCN5A mRNA expression compared with WT (**** p < 0.0001). Data were normalized to WT. ( B ) Quantification of intracellular SCN5A plasmid DNA levels. No significant differences were observed among WT, WT+MUT, and MUT groups (ns), indicating comparable transfection efficiency. ( C ) SCN5A mRNA decay following transcriptional inhibition with actinomycin D. Relative mRNA remaining was normalized to the 0-h time point. The D372H group exhibited accelerated mRNA decay compared with WT, indicating reduced transcript stability. ( D ) Representative Western blot images showing total Nav1.5 protein expression in BLANK, CONTROL, WT, WT+MUT, and MUT groups. Molecular weight markers are indicated in kilodaltons (kDa). The predicted molecular weight of native Nav1.5 is approximately 227 kDa; however, in this study SCN5A was expressed as a GFP-tagged fusion protein, resulting in an apparent band migrating above 245 kDa. Tubulin (50 kDa) was used as a loading control. ( E ) Quantitative analysis of Nav1.5 protein expression normalized to Tubulin. Protein levels were significantly reduced in the D372H group compared with WT (* p < 0.05). The WT+MUT group also showed decreased expression compared with WT (** p < 0.01). Data are presented as mean ± SD from at least three independent experiments.

Article Snippet: The membrane was then incubated with primary antibodies against Nav1.5 (1:1000, Cat# 23016-1-AP, Proteintech, Wuhan, China) and Tubulin (1:000, Cat# A03989-1, Boster, Wuhan, China) to detect the target protein and serve as a loading control, respectively.

Techniques: Variant Assay, Expressing, Plasmid Preparation, Quantitative RT-PCR, Transfection, Inhibition, Western Blot, Control, Molecular Weight