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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, 2021-09
86/100 stars

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1) Product Images from "Rad53- and Chk1-Dependent DNA Damage Response Pathways Cooperatively Promote Fungal Pathogenesis and Modulate Antifungal Drug Susceptibility"

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

Journal: mBio

doi: 10.1128/mBio.01726-18

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]).
Figure Legend 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]).

Techniques Used: Sequencing, Migration, Western Blot, Incubation

Related Articles

Western Blot:

Article Title: Regulation of Candida albicans Interaction with Macrophages through the Activation of HOG Pathway by Genistein
Article Snippet: .. The membrane was stripped and total Hog1 was detected using Hog1 (y-215) sc 9079 rabbit polyclonal IgG (Santa Cruz Biotechnology) and HRP-linked anti-rabbit antibody, and visualized using chemiluminescence (ECL Advance Western Blotting Detection Kit, GE Healthcare, Freiburg, Germany). ..

SDS Page:

Article Title: Intracellular mechanism by which arsenite activates the yeast stress MAPK Hog1
Article Snippet: .. SDS–PAGE and immunoblot analysis Proteins were separated by SDS–PAGE (7.5% gels) followed by immunoblot analysis using mouse monoclonal α-Myc antibody (9E10; Santa Cruz), α-HA (16B12; Covance), α-GST (B-14; Santa Cruz), α-tetra His (Qiagen), or goat polyclonal α-Hog1 (yC-20; Santa Cruz) at a dilution of 1:10,000. ..

Article Title: Intracellular mechanism by which arsenite activates the yeast stress MAPK Hog1
Article Snippet: .. Proteins were separated by SDS–PAGE (7.5% gels) followed by immunoblot analysis using mouse monoclonal α-Myc antibody (9E10; Santa Cruz), α-HA (16B12; Covance), α-GST (B-14; Santa Cruz), α-tetra His (Qiagen), or goat polyclonal α-Hog1 (yC-20; Santa Cruz) at a dilution of 1:10,000. ..

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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, 2021-09
    86/100 stars
      Buy from Supplier

    86
    Santa Cruz Biotechnology hog1 specific rabbit polyclonal antibody
    CAM inhibited the phosphorylation of <t>Hog1</t> and mRNA expression of some virulence genes of Cryptococcus associated with capsular formation in C. gattii R265. (A) C. gattii strain R265 was grown to the mid-exponential phase and treated with CAM in liquid YPD medium, and total protein extracts were prepared for Western blot analysis. The same blot was stripped and probed with <t>polyclonal</t> anti-Hog1 antibody as a loading control (Hog1). (B) mRNA expression of LAC1 , LAC2 , CAP59 , and Mpk1 of R265 with CAM (black bar) or without CAM (white bar) exposure for 2 h. The fold change data were calculated relative to the cells without CAM and were normalized to the expression of the internal control, GPD1 . Data are presented as means ± SEM of results from three experiments (*,
    Hog1 Specific Rabbit Polyclonal 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/hog1 specific rabbit polyclonal antibody/product/Santa Cruz Biotechnology
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    hog1 specific rabbit polyclonal antibody - by Bioz Stars, 2021-09
    86/100 stars
      Buy from Supplier

    96
    Santa Cruz Biotechnology hog1 y 215 sc 9079 rabbit polyclonal igg
    ( a ) Phosphorylation of Hog1p of C. albicans strains after treatment with genistein and solvent for 15min. Hog1p was detected by Hog1 (y-215) <t>sc</t> 9079 rabbit <t>polyclonal</t> <t>IgG</t> and phosphorylated Hog1p (Hog1-P) by Phospho-p38 MAPK (Thr180/182) 3D7 rabbit mAb at a site corresponding to 50 kDa; ( b , c ) Effect of genistein on glycerol and ethanol production in C. albicans . ** p
    Hog1 Y 215 Sc 9079 Rabbit Polyclonal Igg, supplied by Santa Cruz Biotechnology, 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/result/hog1 y 215 sc 9079 rabbit polyclonal igg/product/Santa Cruz Biotechnology
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    Price from $9.99 to $1999.99
    hog1 y 215 sc 9079 rabbit polyclonal igg - by Bioz Stars, 2021-09
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    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

    CAM inhibited the phosphorylation of Hog1 and mRNA expression of some virulence genes of Cryptococcus associated with capsular formation in C. gattii R265. (A) C. gattii strain R265 was grown to the mid-exponential phase and treated with CAM in liquid YPD medium, and total protein extracts were prepared for Western blot analysis. The same blot was stripped and probed with polyclonal anti-Hog1 antibody as a loading control (Hog1). (B) mRNA expression of LAC1 , LAC2 , CAP59 , and Mpk1 of R265 with CAM (black bar) or without CAM (white bar) exposure for 2 h. The fold change data were calculated relative to the cells without CAM and were normalized to the expression of the internal control, GPD1 . Data are presented as means ± SEM of results from three experiments (*,

    Journal: Antimicrobial Agents and Chemotherapy

    Article Title: Macrolides Inhibit Capsule Formation of Highly Virulent Cryptococcus gattii and Promote Innate Immune Susceptibility

    doi: 10.1128/AAC.02364-18

    Figure Lengend Snippet: CAM inhibited the phosphorylation of Hog1 and mRNA expression of some virulence genes of Cryptococcus associated with capsular formation in C. gattii R265. (A) C. gattii strain R265 was grown to the mid-exponential phase and treated with CAM in liquid YPD medium, and total protein extracts were prepared for Western blot analysis. The same blot was stripped and probed with polyclonal anti-Hog1 antibody as a loading control (Hog1). (B) mRNA expression of LAC1 , LAC2 , CAP59 , and Mpk1 of R265 with CAM (black bar) or without CAM (white bar) exposure for 2 h. The fold change data were calculated relative to the cells without CAM and were normalized to the expression of the internal control, GPD1 . Data are presented as means ± SEM of results from three experiments (*,

    Article Snippet: After blocking was performed, the membranes were probed with phospho-p38 MAPK (Thr180/Tyr182) rabbit monoclonal antibody (Cell Signaling Technology, catalog no. 4511S) (1:10,000 dilution) and Hog1-specific rabbit polyclonal antibody (Santa Cruz Biotechnology, Inc., Dallas, TX, USA, catalog no. 25757) (1:4,000) by incubation for 1 h at room temperature for phosphorylated Hog1 determination and total Hog1 protein-independent phosphorylation.

    Techniques: Chick Chorioallantoic Membrane Assay, Expressing, Western Blot

    ( a ) Phosphorylation of Hog1p of C. albicans strains after treatment with genistein and solvent for 15min. Hog1p was detected by Hog1 (y-215) sc 9079 rabbit polyclonal IgG and phosphorylated Hog1p (Hog1-P) by Phospho-p38 MAPK (Thr180/182) 3D7 rabbit mAb at a site corresponding to 50 kDa; ( b , c ) Effect of genistein on glycerol and ethanol production in C. albicans . ** p

    Journal: Molecules

    Article Title: Regulation of Candida albicans Interaction with Macrophages through the Activation of HOG Pathway by Genistein

    doi: 10.3390/molecules21020162

    Figure Lengend Snippet: ( a ) Phosphorylation of Hog1p of C. albicans strains after treatment with genistein and solvent for 15min. Hog1p was detected by Hog1 (y-215) sc 9079 rabbit polyclonal IgG and phosphorylated Hog1p (Hog1-P) by Phospho-p38 MAPK (Thr180/182) 3D7 rabbit mAb at a site corresponding to 50 kDa; ( b , c ) Effect of genistein on glycerol and ethanol production in C. albicans . ** p

    Article Snippet: The membrane was stripped and total Hog1 was detected using Hog1 (y-215) sc 9079 rabbit polyclonal IgG (Santa Cruz Biotechnology) and HRP-linked anti-rabbit antibody, and visualized using chemiluminescence (ECL Advance Western Blotting Detection Kit, GE Healthcare, Freiburg, Germany).

    Techniques:

    C. neoformans Rad53 is phosphorylated by both Mec1 and Tel1 kinases in response to DNA damage stress. (A) Schematic outline of Rad53 orthologs in fungi. The box with dashed lines represents the FHA domain, and the black box indicates the kinase domain. The gray inverted triangles and black inverted triangles indicate SQ and TQ sites, respectively. The domain of each protein was analyzed using Pfam ( http://pfam.xfam.org/ ). 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 Rad53 is phosphorylated by both Mec1 and Tel1 kinases in response to DNA damage stress. (A) Schematic outline of Rad53 orthologs in fungi. The box with dashed lines represents the FHA domain, and the black box indicates the kinase domain. The gray inverted triangles and black inverted triangles indicate SQ and TQ sites, respectively. The domain of each protein was analyzed using Pfam ( http://pfam.xfam.org/ ). 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