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    Name:
    Nuclear Fast Red
    Description:
    Nuclear Fast Red is used as a biological stain and is suitable for the determination of calcium Nuclear Fast Red targets nucleic acids and works more quickly than Hematoxylin sc 24973
    Catalog Number:
    SC-215592A
    Price:
    None
    Category:
    Chemicals Stains Dyes Probes Labels Nucleus and Nucleolus Staining Nuclear Fast Red
    Applications:
    Nuclear Fast Red is a fast-acting biological stain that targets nucleic acids
    Purity:
    ≥92%
    Molecular Weight:
    357.27
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    Structured Review

    Santa Cruz Biotechnology hog1
    The <t>Hog1</t> signaling profile is insufficient to predict downstream output
    Nuclear Fast Red is used as a biological stain and is suitable for the determination of calcium Nuclear Fast Red targets nucleic acids and works more quickly than Hematoxylin sc 24973
    https://www.bioz.com/result/hog1/product/Santa Cruz Biotechnology
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    hog1 - by Bioz Stars, 2021-07
    93/100 stars

    Images

    1) Product Images from "MAPK feedback encodes a switch and timer for tunable stress adaptation in yeast"

    Article Title: MAPK feedback encodes a switch and timer for tunable stress adaptation in yeast

    Journal: Science signaling

    doi: 10.1126/scisignal.2005774

    The Hog1 signaling profile is insufficient to predict downstream output
    Figure Legend Snippet: The Hog1 signaling profile is insufficient to predict downstream output

    Techniques Used:

    The Hog1 signaling profile is a linear transducer that converts dose to duration
    Figure Legend Snippet: The Hog1 signaling profile is a linear transducer that converts dose to duration

    Techniques Used:

    Modeling positive and negative feedback by Hog1
    Figure Legend Snippet: Modeling positive and negative feedback by Hog1

    Techniques Used:

    Hog1 encodes dose-to-duration signaling through graded phosphorylation
    Figure Legend Snippet: Hog1 encodes dose-to-duration signaling through graded phosphorylation

    Techniques Used:

    Hog1 executes a tiered adaptive protein induction program over time
    Figure Legend Snippet: Hog1 executes a tiered adaptive protein induction program over time

    Techniques Used:

    The Hog1 signaling profile can be reengineered through component gene deletions
    Figure Legend Snippet: The Hog1 signaling profile can be reengineered through component gene deletions

    Techniques Used:

    Related Articles

    Immunohistochemistry:

    Article Title: miR-221 silencing blocks hepatocellular carcinoma and promotes survival
    Article Snippet: In situ hybridization was performed on the FFPE sections of tumor tissue as described ( ) and were blinded to the experimental conditions. .. Nuclear fast red was used as the counterstain. p27Kip1 , Ki-67, and cleaved caspase-3 (Santa Cruz Biotechnology) were detected using standard immunohistochemistry techniques. .. Co-localization analysis was performed with the Nuance system from Cambridge Research Institute (Woburn, MA) according to their specifications.

    Staining:

    Article Title: DYSFUNCTIONAL AND PRO-INFLAMMATORY REGULATORY T-LYMPHOCYTES ARE ESSENTIAL FOR ADVERSE CARDIAC REMODELING IN ISCHEMIC CARDIOMYOPATHY
    Article Snippet: However, presumably due to low lymphocytic TNFR1 expression, prior studies have failed to detect TNFR1 on Tregs using fluorochrome-conjugated antibodies., To improve flow cytometric TNFR1 detection, we used a primary rabbit anti-mouse TNFR1 antibody and a secondary Alexa-fluor 405-conjugated goat anti-rabbit antibody. .. We stained splenocytes from HF mice for CD4, Foxp3, TNFα and TNFR1, and nuclei with DAPI, and visualized stained cells using imaging flow cytometry (ImageStream). ..

    Article Title: Stem cells derived from burned skin - The future of burn care
    Article Snippet: Slides were incubated in 1% alcian blue 3GX (Santa Cruz Biotechnology) in 3% acetic acid in water for 30 min at RT. .. The stain was washed with tap water then distilled water then counterstained with 0.1% nuclear fast red (Santa Cruz Biotechnology). ..

    Mouse Assay:

    Article Title: DYSFUNCTIONAL AND PRO-INFLAMMATORY REGULATORY T-LYMPHOCYTES ARE ESSENTIAL FOR ADVERSE CARDIAC REMODELING IN ISCHEMIC CARDIOMYOPATHY
    Article Snippet: However, presumably due to low lymphocytic TNFR1 expression, prior studies have failed to detect TNFR1 on Tregs using fluorochrome-conjugated antibodies., To improve flow cytometric TNFR1 detection, we used a primary rabbit anti-mouse TNFR1 antibody and a secondary Alexa-fluor 405-conjugated goat anti-rabbit antibody. .. We stained splenocytes from HF mice for CD4, Foxp3, TNFα and TNFR1, and nuclei with DAPI, and visualized stained cells using imaging flow cytometry (ImageStream). ..

    Imaging:

    Article Title: DYSFUNCTIONAL AND PRO-INFLAMMATORY REGULATORY T-LYMPHOCYTES ARE ESSENTIAL FOR ADVERSE CARDIAC REMODELING IN ISCHEMIC CARDIOMYOPATHY
    Article Snippet: However, presumably due to low lymphocytic TNFR1 expression, prior studies have failed to detect TNFR1 on Tregs using fluorochrome-conjugated antibodies., To improve flow cytometric TNFR1 detection, we used a primary rabbit anti-mouse TNFR1 antibody and a secondary Alexa-fluor 405-conjugated goat anti-rabbit antibody. .. We stained splenocytes from HF mice for CD4, Foxp3, TNFα and TNFR1, and nuclei with DAPI, and visualized stained cells using imaging flow cytometry (ImageStream). ..

    Flow Cytometry:

    Article Title: DYSFUNCTIONAL AND PRO-INFLAMMATORY REGULATORY T-LYMPHOCYTES ARE ESSENTIAL FOR ADVERSE CARDIAC REMODELING IN ISCHEMIC CARDIOMYOPATHY
    Article Snippet: However, presumably due to low lymphocytic TNFR1 expression, prior studies have failed to detect TNFR1 on Tregs using fluorochrome-conjugated antibodies., To improve flow cytometric TNFR1 detection, we used a primary rabbit anti-mouse TNFR1 antibody and a secondary Alexa-fluor 405-conjugated goat anti-rabbit antibody. .. We stained splenocytes from HF mice for CD4, Foxp3, TNFα and TNFR1, and nuclei with DAPI, and visualized stained cells using imaging flow cytometry (ImageStream). ..

    Cytometry:

    Article Title: DYSFUNCTIONAL AND PRO-INFLAMMATORY REGULATORY T-LYMPHOCYTES ARE ESSENTIAL FOR ADVERSE CARDIAC REMODELING IN ISCHEMIC CARDIOMYOPATHY
    Article Snippet: However, presumably due to low lymphocytic TNFR1 expression, prior studies have failed to detect TNFR1 on Tregs using fluorochrome-conjugated antibodies., To improve flow cytometric TNFR1 detection, we used a primary rabbit anti-mouse TNFR1 antibody and a secondary Alexa-fluor 405-conjugated goat anti-rabbit antibody. .. We stained splenocytes from HF mice for CD4, Foxp3, TNFα and TNFR1, and nuclei with DAPI, and visualized stained cells using imaging flow cytometry (ImageStream). ..

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  • 93
    Santa Cruz Biotechnology hog1
    The <t>Hog1</t> signaling profile is insufficient to predict downstream output
    Hog1, supplied by Santa Cruz Biotechnology, 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/result/hog1/product/Santa Cruz Biotechnology
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    hog1 - by Bioz Stars, 2021-07
    93/100 stars
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    93
    Santa Cruz Biotechnology anti hog1 antibody
    Ptc2 inactivates <t>Hog1</t> in vitro. (A) Activated GST-Hog1 was isolated from osmotic stressed yeast and was untreated or treated with Ptc2. Approximately 0.5 μg of GST-Hog1 bound to resin was incubated with 0 to 1.2 μg of Ptc2 for 30 min at 30°C. The resin was washed extensively to remove Ptc2, and GST-Hog1 was incubated with MBP and [γ- 32 P]ATP to assess Hog1 kinase activity. Radiolabel incorporated into MBP was examined by PhosphorImager analysis. (B) Ptc2 inactivates Hog1 by dephosphorylating the phosphothreonine residue (pT) in the activation loop. GST-Hog1 was phosphorylated in vitro, and the sample was untreated (left) or incubated with Ptc2 (right) prior to phosphoamino acid analysis. The radiolabeled amino acids were detected with the PhosphorImager. Arrows, positions of phosphoamino acid standards as revealed by ninhydrin staining.
    Anti Hog1 Antibody, supplied by Santa Cruz Biotechnology, 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/result/anti hog1 antibody/product/Santa Cruz Biotechnology
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti hog1 antibody - by Bioz Stars, 2021-07
    93/100 stars
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    86
    Santa Cruz Biotechnology rabbit polyclonal hog1 antibody
    C. neoformans ). Each protein sequence was retrieved from the genome database and NCBI [ S. cerevisiae , Rad53; C. albicans , Rad53; C. neoformans , Rad53; and S. pombe , Cds1] aa, amino acids. (B and C) Phosphorylation of Rad53 was monitored by analysis of the reduced electrophoretic migration using western blotting with anti-FLAG antibody. The Rad53-4xFLAG strain was grown to the mid-logarithmic phase and then treated with MMS (0.02%) for 2 h. The cell extract was incubated at 30°C for 1 h with or without λ-phosphatase (PPase) and PPase inhibitor (B). Rad53 was phosphorylated in response to MMS (0.02%), 4-NQO (0.15 µg/ml), and bleomycin (3 µg/ml) (C). (D) Both Tel1 and Mec1 regulate Rad53 phosphorylation in response to DNA damage stress. WT Rad53-4xFLAG, mec1 Δ Rad53-4xFLAG, tel1 Δ Rad53-4xFLAG, and mec1 Δ tel1 Δ Rad53-4xFLAG strains were treated with MMS (0.02%), and then total protein was extracted from each strain for immunoblot analysis. Rad53 phosphorylation levels were monitored using anti-FLAG antibody. The same blot was stripped and then reprobed with <t>polyclonal</t> <t>anti-Hog1</t> antibody for the loading control. (E) Mec1 and Tel1 play redundant roles in DNA damage stress response in C. neoformans . (Strains: WT Rad53-4xFLAG [YSB3806], Rad53-4xFLAG mec1 Δ [KW102], Rad53-4xFLAG tel1 Δ [KW104], Rad53-4xFLAG mec1 Δ tel1 Δ [KW449], mec1 Δ [YSB3611], tel1 Δ [YSB3844], mec1 Δ tel1 Δ [KW480], rad53 Δ [YSB3785], rad53 Δ+ RAD53 [KW1], and tel1 Δ rad53 Δ [KW106]).
    Rabbit Polyclonal Hog1 Antibody, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit polyclonal hog1 antibody/product/Santa Cruz Biotechnology
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit polyclonal hog1 antibody - by Bioz Stars, 2021-07
    86/100 stars
      Buy from Supplier

    Image Search Results


    The Hog1 signaling profile is insufficient to predict downstream output

    Journal: Science signaling

    Article Title: MAPK feedback encodes a switch and timer for tunable stress adaptation in yeast

    doi: 10.1126/scisignal.2005774

    Figure Lengend Snippet: The Hog1 signaling profile is insufficient to predict downstream output

    Article Snippet: Once activated, Hog1 translocates to the nucleus at a rate kn and trans-locates out of the nucleus at a rate kc .

    Techniques:

    The Hog1 signaling profile is a linear transducer that converts dose to duration

    Journal: Science signaling

    Article Title: MAPK feedback encodes a switch and timer for tunable stress adaptation in yeast

    doi: 10.1126/scisignal.2005774

    Figure Lengend Snippet: The Hog1 signaling profile is a linear transducer that converts dose to duration

    Article Snippet: Once activated, Hog1 translocates to the nucleus at a rate kn and trans-locates out of the nucleus at a rate kc .

    Techniques:

    Modeling positive and negative feedback by Hog1

    Journal: Science signaling

    Article Title: MAPK feedback encodes a switch and timer for tunable stress adaptation in yeast

    doi: 10.1126/scisignal.2005774

    Figure Lengend Snippet: Modeling positive and negative feedback by Hog1

    Article Snippet: Once activated, Hog1 translocates to the nucleus at a rate kn and trans-locates out of the nucleus at a rate kc .

    Techniques:

    Hog1 encodes dose-to-duration signaling through graded phosphorylation

    Journal: Science signaling

    Article Title: MAPK feedback encodes a switch and timer for tunable stress adaptation in yeast

    doi: 10.1126/scisignal.2005774

    Figure Lengend Snippet: Hog1 encodes dose-to-duration signaling through graded phosphorylation

    Article Snippet: Once activated, Hog1 translocates to the nucleus at a rate kn and trans-locates out of the nucleus at a rate kc .

    Techniques:

    Hog1 executes a tiered adaptive protein induction program over time

    Journal: Science signaling

    Article Title: MAPK feedback encodes a switch and timer for tunable stress adaptation in yeast

    doi: 10.1126/scisignal.2005774

    Figure Lengend Snippet: Hog1 executes a tiered adaptive protein induction program over time

    Article Snippet: Once activated, Hog1 translocates to the nucleus at a rate kn and trans-locates out of the nucleus at a rate kc .

    Techniques:

    The Hog1 signaling profile can be reengineered through component gene deletions

    Journal: Science signaling

    Article Title: MAPK feedback encodes a switch and timer for tunable stress adaptation in yeast

    doi: 10.1126/scisignal.2005774

    Figure Lengend Snippet: The Hog1 signaling profile can be reengineered through component gene deletions

    Article Snippet: Once activated, Hog1 translocates to the nucleus at a rate kn and trans-locates out of the nucleus at a rate kc .

    Techniques:

    Ptc2 inactivates Hog1 in vitro. (A) Activated GST-Hog1 was isolated from osmotic stressed yeast and was untreated or treated with Ptc2. Approximately 0.5 μg of GST-Hog1 bound to resin was incubated with 0 to 1.2 μg of Ptc2 for 30 min at 30°C. The resin was washed extensively to remove Ptc2, and GST-Hog1 was incubated with MBP and [γ- 32 P]ATP to assess Hog1 kinase activity. Radiolabel incorporated into MBP was examined by PhosphorImager analysis. (B) Ptc2 inactivates Hog1 by dephosphorylating the phosphothreonine residue (pT) in the activation loop. GST-Hog1 was phosphorylated in vitro, and the sample was untreated (left) or incubated with Ptc2 (right) prior to phosphoamino acid analysis. The radiolabeled amino acids were detected with the PhosphorImager. Arrows, positions of phosphoamino acid standards as revealed by ninhydrin staining.

    Journal: Eukaryotic Cell

    Article Title: Role of Ptc2 Type 2C Ser/Thr Phosphatase in Yeast High-Osmolarity Glycerol Pathway Inactivation

    doi: 10.1128/EC.1.6.1032-1040.2002

    Figure Lengend Snippet: Ptc2 inactivates Hog1 in vitro. (A) Activated GST-Hog1 was isolated from osmotic stressed yeast and was untreated or treated with Ptc2. Approximately 0.5 μg of GST-Hog1 bound to resin was incubated with 0 to 1.2 μg of Ptc2 for 30 min at 30°C. The resin was washed extensively to remove Ptc2, and GST-Hog1 was incubated with MBP and [γ- 32 P]ATP to assess Hog1 kinase activity. Radiolabel incorporated into MBP was examined by PhosphorImager analysis. (B) Ptc2 inactivates Hog1 by dephosphorylating the phosphothreonine residue (pT) in the activation loop. GST-Hog1 was phosphorylated in vitro, and the sample was untreated (left) or incubated with Ptc2 (right) prior to phosphoamino acid analysis. The radiolabeled amino acids were detected with the PhosphorImager. Arrows, positions of phosphoamino acid standards as revealed by ninhydrin staining.

    Article Snippet: GST-Ptc2 and GST were detected with an anti-GST antibody (Pharmacia), and Hog1 was visualized with an anti-Hog1 antibody (yC-20; Santa Cruz Biotechnology).

    Techniques: In Vitro, Isolation, Incubation, Activity Assay, Activation Assay, Phosphoamino Acid Analysis, Staining

    Ptc2 inactivates Hog1 in vivo. (A) Overexpression of PTC2 inhibits osmotic-stress-induced Hog1 activation. Hog1 kinase activity in PTC2 overexpressor IMY105, carrying pKT-PTC2 and pHOG1-ha2, and in the control strain, carrying empty vector pKT and pHOG1-ha2, was examined. Before (time zero) and after exposure to osmotic stress (0.4 M NaCl) for various times, Hog1-HA was immunoprecipitated and incubated with MBP and [γ- 32 P]ATP. Radiolabel incorporated into MBP was examined with the PhosphorImager. The graph shows the means of three independent experiments ± standard errors of the means (SEM). (B) Hog1 is hyperactivated in a strain lacking PTC2 and PTC3 . Hog1 kinase activity was monitored prior to and following osmotic stress in a hog1 Δ strain (IMY100) and in a ptc2 Δ ptc3 Δ hog1 Δ strain (CAY9), each carrying a Hog1-HA-expressing plasmid. Hog1 kinase activity was monitored as described for panel A. The graph shows the means of six independent experiments ± SEM. (C) Ptc2 does not inactivate Pbs2 in vivo. Hog1-pY in the 334 strain carrying a plasmid overexpressing PTC2 or an empty vector was examined. The level of Hog1-pY prior to and following osmotic stress was monitored by immunoblotting with an anti-pY antibody. Total Hog1 protein was examined by blotting with an anti-Hog1 antibody.

    Journal: Eukaryotic Cell

    Article Title: Role of Ptc2 Type 2C Ser/Thr Phosphatase in Yeast High-Osmolarity Glycerol Pathway Inactivation

    doi: 10.1128/EC.1.6.1032-1040.2002

    Figure Lengend Snippet: Ptc2 inactivates Hog1 in vivo. (A) Overexpression of PTC2 inhibits osmotic-stress-induced Hog1 activation. Hog1 kinase activity in PTC2 overexpressor IMY105, carrying pKT-PTC2 and pHOG1-ha2, and in the control strain, carrying empty vector pKT and pHOG1-ha2, was examined. Before (time zero) and after exposure to osmotic stress (0.4 M NaCl) for various times, Hog1-HA was immunoprecipitated and incubated with MBP and [γ- 32 P]ATP. Radiolabel incorporated into MBP was examined with the PhosphorImager. The graph shows the means of three independent experiments ± standard errors of the means (SEM). (B) Hog1 is hyperactivated in a strain lacking PTC2 and PTC3 . Hog1 kinase activity was monitored prior to and following osmotic stress in a hog1 Δ strain (IMY100) and in a ptc2 Δ ptc3 Δ hog1 Δ strain (CAY9), each carrying a Hog1-HA-expressing plasmid. Hog1 kinase activity was monitored as described for panel A. The graph shows the means of six independent experiments ± SEM. (C) Ptc2 does not inactivate Pbs2 in vivo. Hog1-pY in the 334 strain carrying a plasmid overexpressing PTC2 or an empty vector was examined. The level of Hog1-pY prior to and following osmotic stress was monitored by immunoblotting with an anti-pY antibody. Total Hog1 protein was examined by blotting with an anti-Hog1 antibody.

    Article Snippet: GST-Ptc2 and GST were detected with an anti-GST antibody (Pharmacia), and Hog1 was visualized with an anti-Hog1 antibody (yC-20; Santa Cruz Biotechnology).

    Techniques: In Vivo, Over Expression, Activation Assay, Activity Assay, Plasmid Preparation, Immunoprecipitation, Incubation, Expressing

    The temperature sensitivity of strains lacking PTC s and PTP2 is due to HOG1. (A) Strains lacking PTC s and PTP2 exhibit growth defects at 37°C. The growth of ptp2 Δ (IMY21a), ptc2 Δ ptc3 Δ (IMY124), ptc3 Δ ptp2 Δ (IMY127b), ptc2 Δ ptc3 Δ ptp2 Δ (IMY128), and wild-type (BBY45) strains on rich medium (yeast extract-peptone-dextrose [YPD]) at 37 and 30°C was compared. (B) The growth defects of the ptc3 Δ ptp2 Δ and ptc2 Δ ptc3 Δ ptp2 Δ strains at 37°C were suppressed by deleting HOG1 . The strains lacking HOG1 were JMY1 ( ptc3 Δ ptp2 Δ hog1 Δ and JMY2 ( ptc2 Δ ptc3 Δ ptp2 Δ hog1 Δ) and were compared on YPD at 37 and 30°C.

    Journal: Eukaryotic Cell

    Article Title: Role of Ptc2 Type 2C Ser/Thr Phosphatase in Yeast High-Osmolarity Glycerol Pathway Inactivation

    doi: 10.1128/EC.1.6.1032-1040.2002

    Figure Lengend Snippet: The temperature sensitivity of strains lacking PTC s and PTP2 is due to HOG1. (A) Strains lacking PTC s and PTP2 exhibit growth defects at 37°C. The growth of ptp2 Δ (IMY21a), ptc2 Δ ptc3 Δ (IMY124), ptc3 Δ ptp2 Δ (IMY127b), ptc2 Δ ptc3 Δ ptp2 Δ (IMY128), and wild-type (BBY45) strains on rich medium (yeast extract-peptone-dextrose [YPD]) at 37 and 30°C was compared. (B) The growth defects of the ptc3 Δ ptp2 Δ and ptc2 Δ ptc3 Δ ptp2 Δ strains at 37°C were suppressed by deleting HOG1 . The strains lacking HOG1 were JMY1 ( ptc3 Δ ptp2 Δ hog1 Δ and JMY2 ( ptc2 Δ ptc3 Δ ptp2 Δ hog1 Δ) and were compared on YPD at 37 and 30°C.

    Article Snippet: GST-Ptc2 and GST were detected with an anti-GST antibody (Pharmacia), and Hog1 was visualized with an anti-Hog1 antibody (yC-20; Santa Cruz Biotechnology).

    Techniques:

    Model of the yeast hyperosmotic-response MAPK pathway. (A) The osmotic signals from Sho1 or Sln1 are transduced by unique components and converge to activate Pbs2. The Sho1 branch requires Cdc42, Ste20, and Ste50 to activate Ste11. The Sln1 protein activates Ssk2 and Ssk22 through Ypd1 and Ssk1. Any of Ste11, Ssk2, or Ssk22 is able to activate Pbs2, which then phosphorylates Hog1, resulting in translocation of Hog1 to the nucleus that regulates responsible genes expression for osmoadaptation. (B) The schematic diagram of Sho1 and Sln1 orthologs in Botrytis cinerea . The functional domains were retrieved in EnsemblFungi ( https://fungi.ensembl.org/Botrytis_cinerea/Info/Index ). (C) Subcellular localization of BcSho1-GFP and BcSln1-GFP fusion proteins in B. cienrea . Bars, 10 μm.

    Journal: Frontiers in Microbiology

    Article Title: The Sensor Proteins BcSho1 and BcSln1 Are Involved in, Though Not Essential to, Vegetative Differentiation, Pathogenicity and Osmotic Stress Tolerance in Botrytis cinerea

    doi: 10.3389/fmicb.2019.00328

    Figure Lengend Snippet: Model of the yeast hyperosmotic-response MAPK pathway. (A) The osmotic signals from Sho1 or Sln1 are transduced by unique components and converge to activate Pbs2. The Sho1 branch requires Cdc42, Ste20, and Ste50 to activate Ste11. The Sln1 protein activates Ssk2 and Ssk22 through Ypd1 and Ssk1. Any of Ste11, Ssk2, or Ssk22 is able to activate Pbs2, which then phosphorylates Hog1, resulting in translocation of Hog1 to the nucleus that regulates responsible genes expression for osmoadaptation. (B) The schematic diagram of Sho1 and Sln1 orthologs in Botrytis cinerea . The functional domains were retrieved in EnsemblFungi ( https://fungi.ensembl.org/Botrytis_cinerea/Info/Index ). (C) Subcellular localization of BcSho1-GFP and BcSln1-GFP fusion proteins in B. cienrea . Bars, 10 μm.

    Article Snippet: In addition, the samples were detected with anti-Hog1 antibody (C-terminal anti-Hog1) (Santa Cruz Biotechnology, Santa Cruz, CA, United States) as a reference.

    Techniques: Translocation Assay, Expressing, Functional Assay

    C. neoformans ). Each protein sequence was retrieved from the genome database and NCBI [ S. cerevisiae , Rad53; C. albicans , Rad53; C. neoformans , Rad53; and S. pombe , Cds1] aa, amino acids. (B and C) Phosphorylation of Rad53 was monitored by analysis of the reduced electrophoretic migration using western blotting with anti-FLAG antibody. The Rad53-4xFLAG strain was grown to the mid-logarithmic phase and then treated with MMS (0.02%) for 2 h. The cell extract was incubated at 30°C for 1 h with or without λ-phosphatase (PPase) and PPase inhibitor (B). Rad53 was phosphorylated in response to MMS (0.02%), 4-NQO (0.15 µg/ml), and bleomycin (3 µg/ml) (C). (D) Both Tel1 and Mec1 regulate Rad53 phosphorylation in response to DNA damage stress. WT Rad53-4xFLAG, mec1 Δ Rad53-4xFLAG, tel1 Δ Rad53-4xFLAG, and mec1 Δ tel1 Δ Rad53-4xFLAG strains were treated with MMS (0.02%), and then total protein was extracted from each strain for immunoblot analysis. Rad53 phosphorylation levels were monitored using anti-FLAG antibody. The same blot was stripped and then reprobed with polyclonal anti-Hog1 antibody for the loading control. (E) Mec1 and Tel1 play redundant roles in DNA damage stress response in C. neoformans . (Strains: WT Rad53-4xFLAG [YSB3806], Rad53-4xFLAG mec1 Δ [KW102], Rad53-4xFLAG tel1 Δ [KW104], Rad53-4xFLAG mec1 Δ tel1 Δ [KW449], mec1 Δ [YSB3611], tel1 Δ [YSB3844], mec1 Δ tel1 Δ [KW480], rad53 Δ [YSB3785], rad53 Δ+ RAD53 [KW1], and tel1 Δ rad53 Δ [KW106]).

    Journal: mBio

    Article Title: Rad53- and Chk1-Dependent DNA Damage Response Pathways Cooperatively Promote Fungal Pathogenesis and Modulate Antifungal Drug Susceptibility

    doi: 10.1128/mBio.01726-18

    Figure Lengend Snippet: C. neoformans ). Each protein sequence was retrieved from the genome database and NCBI [ S. cerevisiae , Rad53; C. albicans , Rad53; C. neoformans , Rad53; and S. pombe , Cds1] aa, amino acids. (B and C) Phosphorylation of Rad53 was monitored by analysis of the reduced electrophoretic migration using western blotting with anti-FLAG antibody. The Rad53-4xFLAG strain was grown to the mid-logarithmic phase and then treated with MMS (0.02%) for 2 h. The cell extract was incubated at 30°C for 1 h with or without λ-phosphatase (PPase) and PPase inhibitor (B). Rad53 was phosphorylated in response to MMS (0.02%), 4-NQO (0.15 µg/ml), and bleomycin (3 µg/ml) (C). (D) Both Tel1 and Mec1 regulate Rad53 phosphorylation in response to DNA damage stress. WT Rad53-4xFLAG, mec1 Δ Rad53-4xFLAG, tel1 Δ Rad53-4xFLAG, and mec1 Δ tel1 Δ Rad53-4xFLAG strains were treated with MMS (0.02%), and then total protein was extracted from each strain for immunoblot analysis. Rad53 phosphorylation levels were monitored using anti-FLAG antibody. The same blot was stripped and then reprobed with polyclonal anti-Hog1 antibody for the loading control. (E) Mec1 and Tel1 play redundant roles in DNA damage stress response in C. neoformans . (Strains: WT Rad53-4xFLAG [YSB3806], Rad53-4xFLAG mec1 Δ [KW102], Rad53-4xFLAG tel1 Δ [KW104], Rad53-4xFLAG mec1 Δ tel1 Δ [KW449], mec1 Δ [YSB3611], tel1 Δ [YSB3844], mec1 Δ tel1 Δ [KW480], rad53 Δ [YSB3785], rad53 Δ+ RAD53 [KW1], and tel1 Δ rad53 Δ [KW106]).

    Article Snippet: To monitor Hog1 protein levels as the loading control, a primary rabbit polyclonal Hog1 antibody (SC-9079; Santa Cruz Biotechnology) and a secondary anti-rabbit IgG horseradish peroxidase-conjugated antibody (A6154; Sigma) were used.

    Techniques: Sequencing, Migration, Western Blot, Incubation