solid phase resins  (Chem Impex International)

Journal: International Journal of Molecular Sciences

Article Title: Regulation of Disease-Resistance Genes against CWMV Infection by NbHAG1-Mediated H3K36ac

doi: 10.3390/ijms25052800

Figure Lengend Snippet: NbHAG1 mediates H3K36ac to regulate downstream gene expression. ( A ) Western blot detection of the protein content of H3ac in NbHAG1 knockout mutants, WT and NbHAG1 overexpression mutant plants. The protein content in the different samples was then determined by Ponceau S. Viral proteins were quantified using ImageJ software. ( B ) The histone H3 acetylation site of NbHAG1 was identified by LC-MS/MS, and the acetylation site is shown in red. ( C – E ) LC-MS/MS spectra of histone H3 acetylated peptides of CRISPER NbHAG1 and OENbHAG1 (*, p < 0.05). ( F ) Western blot detection of the protein content of H3K36ac in NbHAG1 knockout mutants, WT and NbHAG1 overexpression mutant plants. The protein content in the different samples was then determined by Ponceau S. Viral proteins were quantified using ImageJ software.

Article Snippet: Antibodies used in this study for CUT&Tag were H3K36ac (39379, Active Motif, California, USA), IgG or rabbit IgG (10500C, Invitrogen, Waltham, USA).

Techniques: Gene Expression, Western Blot, Knock-Out, Over Expression, Mutagenesis, Software, Liquid Chromatography with Mass Spectroscopy

Journal: International Journal of Molecular Sciences

Article Title: Regulation of Disease-Resistance Genes against CWMV Infection by NbHAG1-Mediated H3K36ac

doi: 10.3390/ijms25052800

Figure Lengend Snippet: NbHAG1 mediates H3K36ac to regulate downstream gene expression. ( A ) Schematic representation of the fragment between 500 bp before and 500 bp after the downstream gene TSS. The fragments between 500 bp before and after the downstream gene TSS were divided into 4 small fragments, named 1–4 fragments. ( B – F ) Cut&Tag-qPCR to analyze the enrichment of H3K36 marks at the fragment between 500 bp before and 500 bp after the downstream gene TSS in NbHAG1 overexpressing wheat plants. The enrichment levels were compared with DNA spike-in for enrichment detection. Data presented are the mean ± SD of three biological samples per treatment. Each biological sample had three technical replicates. Significant differences between treatments were determined using Student’s t test (*, p < 0.05).

Article Snippet: Antibodies used in this study for CUT&Tag were H3K36ac (39379, Active Motif, California, USA), IgG or rabbit IgG (10500C, Invitrogen, Waltham, USA).

Techniques: Gene Expression

Journal: International Journal of Molecular Sciences

Article Title: Regulation of Disease-Resistance Genes against CWMV Infection by NbHAG1-Mediated H3K36ac

doi: 10.3390/ijms25052800

Figure Lengend Snippet: Working model showing the regulatory role of histone acetylase NbHAG1 in the activation of disease resistance genes by modulating histone H3 lysine 36 acetylation (H3K36ac). Following CWMV infection, there is a notable up-regulation in NbHAG1 expression. As an acetylase, NbHAG1 significantly enhances the acetylation status of H3K36ac, thereby promoting the activation of downstream genes such as NbERF109 and NbMKK2 . This activation cascade effectively suppresses CWMV infection.

Article Snippet: Antibodies used in this study for CUT&Tag were H3K36ac (39379, Active Motif, California, USA), IgG or rabbit IgG (10500C, Invitrogen, Waltham, USA).

Techniques: Activation Assay, Infection, Expressing

Journal: International Journal of Molecular Sciences

Article Title: Regulation of Disease-Resistance Genes against CWMV Infection by NbHAG1-Mediated H3K36ac

doi: 10.3390/ijms25052800

Figure Lengend Snippet: NbHAG1 mediates H3K36ac to regulate downstream gene expression. ( A ) Western blot detection of the protein content of H3ac in NbHAG1 knockout mutants, WT and NbHAG1 overexpression mutant plants. The protein content in the different samples was then determined by Ponceau S. Viral proteins were quantified using ImageJ software. ( B ) The histone H3 acetylation site of NbHAG1 was identified by LC-MS/MS, and the acetylation site is shown in red. ( C – E ) LC-MS/MS spectra of histone H3 acetylated peptides of CRISPER NbHAG1 and OENbHAG1 (*, p < 0.05). ( F ) Western blot detection of the protein content of H3K36ac in NbHAG1 knockout mutants, WT and NbHAG1 overexpression mutant plants. The protein content in the different samples was then determined by Ponceau S. Viral proteins were quantified using ImageJ software.

Article Snippet: The bead-bound protoplasts were incubated in primary antibody buffer containing an anti-H3K36ac antibody (39379, Active Motif, California, USA) at 4 °C by rotating overnight.

Techniques: Gene Expression, Western Blot, Knock-Out, Over Expression, Mutagenesis, Software, Liquid Chromatography with Mass Spectroscopy

Journal: International Journal of Molecular Sciences

Article Title: Regulation of Disease-Resistance Genes against CWMV Infection by NbHAG1-Mediated H3K36ac

doi: 10.3390/ijms25052800

Figure Lengend Snippet: NbHAG1 mediates H3K36ac to regulate downstream gene expression. ( A ) Schematic representation of the fragment between 500 bp before and 500 bp after the downstream gene TSS. The fragments between 500 bp before and after the downstream gene TSS were divided into 4 small fragments, named 1–4 fragments. ( B – F ) Cut&Tag-qPCR to analyze the enrichment of H3K36 marks at the fragment between 500 bp before and 500 bp after the downstream gene TSS in NbHAG1 overexpressing wheat plants. The enrichment levels were compared with DNA spike-in for enrichment detection. Data presented are the mean ± SD of three biological samples per treatment. Each biological sample had three technical replicates. Significant differences between treatments were determined using Student’s t test (*, p < 0.05).

Article Snippet: The bead-bound protoplasts were incubated in primary antibody buffer containing an anti-H3K36ac antibody (39379, Active Motif, California, USA) at 4 °C by rotating overnight.

Techniques: Gene Expression

Journal: International Journal of Molecular Sciences

Article Title: Regulation of Disease-Resistance Genes against CWMV Infection by NbHAG1-Mediated H3K36ac

doi: 10.3390/ijms25052800

Figure Lengend Snippet: Working model showing the regulatory role of histone acetylase NbHAG1 in the activation of disease resistance genes by modulating histone H3 lysine 36 acetylation (H3K36ac). Following CWMV infection, there is a notable up-regulation in NbHAG1 expression. As an acetylase, NbHAG1 significantly enhances the acetylation status of H3K36ac, thereby promoting the activation of downstream genes such as NbERF109 and NbMKK2 . This activation cascade effectively suppresses CWMV infection.

Article Snippet: The bead-bound protoplasts were incubated in primary antibody buffer containing an anti-H3K36ac antibody (39379, Active Motif, California, USA) at 4 °C by rotating overnight.

Techniques: Activation Assay, Infection, Expressing