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Gl Slt2 interacts with Gl Myb. ( A ) Y2H assay detection of the interaction between Gl Slt2 and Gl Myb. pGBKT7-Slt2, <t>pGADT7-Myb,</t> pGADT7-Myb 1–163 , and pGADT7-Myb 164–629 were cotransformed into the Y2H strain. SD-Leu-Trp medium was used for testing successful mating, and SD-Ade-His-Leu-Trp/X-α-gal medium was used for testing interactions. The combination of pGBKT7-53 and pGADT7-T was used as the positive control. ( B ) BiFC verified the interaction between Gl Slt2 and Gl Myb. PVN-Slt2 containing the fragment of Venus-N, PVC-Myb, PVC-Myb 1–163 , and PVC-Myb 164–629 containing the fragment of Venus-C were cotransformed into the SFY2620 yeast strain (scale bar = 100 µm). In addition, the recombinant plasmids PVN and PVC, PVN-Slt2, and PVC, as well as PVN and PVC-Myb, along with their truncated forms (PVC-Myb 1–163 and PVC-Myb 164–629 ), were cotransformed into yeast as negative controls. DIC, yeast cell morphology under the normal white field of view; Venus, a variant of GFP, yeast cell morphology under the green fluorescence. ( C ) Co-IP detection of the interaction between endogenous Gl Slt2 and Gl Myb. Immunoprecipitation of mycelial, collected from the liquid culture medium with microcrystalline cellulose as the sole carbon source, lysates with control rabbit IgG or anti- Gl Slt2 antibody. The IP products were detected by Western blotting with an anti- Gl Slt2 antibody to assess the accuracy of the experiment. In addition, the IP product was probed with an anti- Gl Myb antibody to specifically detect the interaction between Gl Slt2 and endogenous Gl Myb. Whole-cell extracts (Input) show the results of Western blot with anti- Gl Slt2 antibody and anti- Gl Myb antibody as controls.
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Gl Slt2 interacts with Gl Myb. ( A ) Y2H assay detection of the interaction between Gl Slt2 and Gl Myb. pGBKT7-Slt2, <t>pGADT7-Myb,</t> pGADT7-Myb 1–163 , and pGADT7-Myb 164–629 were cotransformed into the Y2H strain. SD-Leu-Trp medium was used for testing successful mating, and SD-Ade-His-Leu-Trp/X-α-gal medium was used for testing interactions. The combination of pGBKT7-53 and pGADT7-T was used as the positive control. ( B ) BiFC verified the interaction between Gl Slt2 and Gl Myb. PVN-Slt2 containing the fragment of Venus-N, PVC-Myb, PVC-Myb 1–163 , and PVC-Myb 164–629 containing the fragment of Venus-C were cotransformed into the SFY2620 yeast strain (scale bar = 100 µm). In addition, the recombinant plasmids PVN and PVC, PVN-Slt2, and PVC, as well as PVN and PVC-Myb, along with their truncated forms (PVC-Myb 1–163 and PVC-Myb 164–629 ), were cotransformed into yeast as negative controls. DIC, yeast cell morphology under the normal white field of view; Venus, a variant of GFP, yeast cell morphology under the green fluorescence. ( C ) Co-IP detection of the interaction between endogenous Gl Slt2 and Gl Myb. Immunoprecipitation of mycelial, collected from the liquid culture medium with microcrystalline cellulose as the sole carbon source, lysates with control rabbit IgG or anti- Gl Slt2 antibody. The IP products were detected by Western blotting with an anti- Gl Slt2 antibody to assess the accuracy of the experiment. In addition, the IP product was probed with an anti- Gl Myb antibody to specifically detect the interaction between Gl Slt2 and endogenous Gl Myb. Whole-cell extracts (Input) show the results of Western blot with anti- Gl Slt2 antibody and anti- Gl Myb antibody as controls.
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Gl Slt2 interacts with Gl Myb. ( A ) Y2H assay detection of the interaction between Gl Slt2 and Gl Myb. pGBKT7-Slt2, <t>pGADT7-Myb,</t> pGADT7-Myb 1–163 , and pGADT7-Myb 164–629 were cotransformed into the Y2H strain. SD-Leu-Trp medium was used for testing successful mating, and SD-Ade-His-Leu-Trp/X-α-gal medium was used for testing interactions. The combination of pGBKT7-53 and pGADT7-T was used as the positive control. ( B ) BiFC verified the interaction between Gl Slt2 and Gl Myb. PVN-Slt2 containing the fragment of Venus-N, PVC-Myb, PVC-Myb 1–163 , and PVC-Myb 164–629 containing the fragment of Venus-C were cotransformed into the SFY2620 yeast strain (scale bar = 100 µm). In addition, the recombinant plasmids PVN and PVC, PVN-Slt2, and PVC, as well as PVN and PVC-Myb, along with their truncated forms (PVC-Myb 1–163 and PVC-Myb 164–629 ), were cotransformed into yeast as negative controls. DIC, yeast cell morphology under the normal white field of view; Venus, a variant of GFP, yeast cell morphology under the green fluorescence. ( C ) Co-IP detection of the interaction between endogenous Gl Slt2 and Gl Myb. Immunoprecipitation of mycelial, collected from the liquid culture medium with microcrystalline cellulose as the sole carbon source, lysates with control rabbit IgG or anti- Gl Slt2 antibody. The IP products were detected by Western blotting with an anti- Gl Slt2 antibody to assess the accuracy of the experiment. In addition, the IP product was probed with an anti- Gl Myb antibody to specifically detect the interaction between Gl Slt2 and endogenous Gl Myb. Whole-cell extracts (Input) show the results of Western blot with anti- Gl Slt2 antibody and anti- Gl Myb antibody as controls.
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Gl Slt2 interacts with Gl Myb. ( A ) Y2H assay detection of the interaction between Gl Slt2 and Gl Myb. pGBKT7-Slt2, pGADT7-Myb, pGADT7-Myb 1–163 , and pGADT7-Myb 164–629 were cotransformed into the Y2H strain. SD-Leu-Trp medium was used for testing successful mating, and SD-Ade-His-Leu-Trp/X-α-gal medium was used for testing interactions. The combination of pGBKT7-53 and pGADT7-T was used as the positive control. ( B ) BiFC verified the interaction between Gl Slt2 and Gl Myb. PVN-Slt2 containing the fragment of Venus-N, PVC-Myb, PVC-Myb 1–163 , and PVC-Myb 164–629 containing the fragment of Venus-C were cotransformed into the SFY2620 yeast strain (scale bar = 100 µm). In addition, the recombinant plasmids PVN and PVC, PVN-Slt2, and PVC, as well as PVN and PVC-Myb, along with their truncated forms (PVC-Myb 1–163 and PVC-Myb 164–629 ), were cotransformed into yeast as negative controls. DIC, yeast cell morphology under the normal white field of view; Venus, a variant of GFP, yeast cell morphology under the green fluorescence. ( C ) Co-IP detection of the interaction between endogenous Gl Slt2 and Gl Myb. Immunoprecipitation of mycelial, collected from the liquid culture medium with microcrystalline cellulose as the sole carbon source, lysates with control rabbit IgG or anti- Gl Slt2 antibody. The IP products were detected by Western blotting with an anti- Gl Slt2 antibody to assess the accuracy of the experiment. In addition, the IP product was probed with an anti- Gl Myb antibody to specifically detect the interaction between Gl Slt2 and endogenous Gl Myb. Whole-cell extracts (Input) show the results of Western blot with anti- Gl Slt2 antibody and anti- Gl Myb antibody as controls.

Journal: mBio

Article Title: Gl Slt2 positively regulates Gl Myb-mediated cellulose utilization in Ganoderma lucidum

doi: 10.1128/mbio.01812-25

Figure Lengend Snippet: Gl Slt2 interacts with Gl Myb. ( A ) Y2H assay detection of the interaction between Gl Slt2 and Gl Myb. pGBKT7-Slt2, pGADT7-Myb, pGADT7-Myb 1–163 , and pGADT7-Myb 164–629 were cotransformed into the Y2H strain. SD-Leu-Trp medium was used for testing successful mating, and SD-Ade-His-Leu-Trp/X-α-gal medium was used for testing interactions. The combination of pGBKT7-53 and pGADT7-T was used as the positive control. ( B ) BiFC verified the interaction between Gl Slt2 and Gl Myb. PVN-Slt2 containing the fragment of Venus-N, PVC-Myb, PVC-Myb 1–163 , and PVC-Myb 164–629 containing the fragment of Venus-C were cotransformed into the SFY2620 yeast strain (scale bar = 100 µm). In addition, the recombinant plasmids PVN and PVC, PVN-Slt2, and PVC, as well as PVN and PVC-Myb, along with their truncated forms (PVC-Myb 1–163 and PVC-Myb 164–629 ), were cotransformed into yeast as negative controls. DIC, yeast cell morphology under the normal white field of view; Venus, a variant of GFP, yeast cell morphology under the green fluorescence. ( C ) Co-IP detection of the interaction between endogenous Gl Slt2 and Gl Myb. Immunoprecipitation of mycelial, collected from the liquid culture medium with microcrystalline cellulose as the sole carbon source, lysates with control rabbit IgG or anti- Gl Slt2 antibody. The IP products were detected by Western blotting with an anti- Gl Slt2 antibody to assess the accuracy of the experiment. In addition, the IP product was probed with an anti- Gl Myb antibody to specifically detect the interaction between Gl Slt2 and endogenous Gl Myb. Whole-cell extracts (Input) show the results of Western blot with anti- Gl Slt2 antibody and anti- Gl Myb antibody as controls.

Article Snippet: The yeast two-hybrid vectors pGADT7 and pGBKT7 were purchased from Clontech (Mountain View, CA, USA).

Techniques: Y2H Assay, Positive Control, Recombinant, Variant Assay, Fluorescence, Co-Immunoprecipitation Assay, Immunoprecipitation, Control, Western Blot