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Image Search Results
Journal: Horticulture Research
Article Title: ALA reverses ABA-induced stomatal closure by modulating PP2AC and SnRK2.6 activity in apple leaves
doi: 10.1093/hr/uhad067
Figure Lengend Snippet: ALA significantly increases abundance and phosphorylation of the MdPP2AC protein. A: ALA promotes MdPP2AC abundance when α-tubulin (actin) was used as loading control. Molecular mass (kDa) markers are shown on the left. All experiments were repeated at least three times with similar results. B: Total plant proteins were extracted and immunoprecipitated with a phosphorylated MdPP2AC antibody. Image J software was used to estimate the relative abundance of total ( C ) and phosphorylated ( D ) MdPP2AC protein. Data are means of three independent biological replicates. The same lowercase letters in C or D indicate no significant difference at p = 0.05.
Article Snippet: The blots were probed with a customized anti-MdPP2AC peptide-raised (
Techniques: Immunoprecipitation, Software
Journal: Horticulture Research
Article Title: ALA reverses ABA-induced stomatal closure by modulating PP2AC and SnRK2.6 activity in apple leaves
doi: 10.1093/hr/uhad067
Figure Lengend Snippet: ALA promotes the interaction of MdPP2AC with other MdPP2A subunits . A: Quadruple dropout medium (SD/−Ade/–His/−Leu/−Trp supplemented with X-α-Gal and AbA) with 0.5 mg L −1 ALA was used in Y2H assays to test potential interactions. BD-53 plus AD-T was applied for the positive control. The β-galactosidase activity was detected by ONPG (o-nitrophenyl β-D-galactopyranoside) assay. B: FLC assays. C: Intensity of the luciferase interaction signal was measured with ImageJ software. Values are means ± SE from three independent experiments ( * p < 0.05).
Article Snippet: The blots were probed with a customized anti-MdPP2AC peptide-raised (
Techniques: Positive Control, Activity Assay, Luciferase, Software
Journal: Horticulture Research
Article Title: ALA reverses ABA-induced stomatal closure by modulating PP2AC and SnRK2.6 activity in apple leaves
doi: 10.1093/hr/uhad067
Figure Lengend Snippet: ALA inhibits the ABA-induced expression of MdSnRK2.6 and promotes MdSnRK2.6 dephosphorylation, and MdSnRK2.6 interacts with MdPP2AC . A: Expression of MdSnRK2.6 in apple leaves under ALA and/or ABA treatment. B: Western blotting assay of phosphorylated MdSnRK2.6, whose abundance was specifically induced by ABA but reduced by ALA. The total proteins of apple leaf epidermis under different treatments were extracted before immunoprecipitation with a specific phosphorylated SnRK2.6 antibody. C: Quantification of the phosphorylated SnRK2.6 levels in B . The same lowercase letters in A or C indicate no significant difference at p = 0.05. D: The subcellular localization of MdPP2AC and MdSnRK2.6 in tobacco leaves. The pCAMBIA1305-mCherry membrane marker and pCAMBIA1300-35S:: GFP , 35S:: MdPP2AC , and 35S:: MdSnRK2.6 recombinant plasmids were transiently co-expressed in 4-week-old N. benthamiana leaves. RFP: Red fluorescent protein. GFP: green fluorescent protein. Scale bar: 25 μm. E: A Y2H assay confirmed the interaction of MdPP2AC with MdSnRK2.6. The combination of BD-53 plus AD-T was used as a positive control, and BD-Lam plus AD-T was used as a negative control. F: The interaction of MdPP2AC and MdSnRK2.6 showed by an FLC assay. G: A BiFC assay showed the interaction of MdPP2AC and MdSnRK2.6. H: Pull-down assay. MdSnRK2.6-His and MdPP2AC-GST proteins were expressed in E. coli for immunoprecipitation. The anti-His and anti-GST antibody were used for detection of MdSnRK2.6-His and MdPP2AC-GST, respectively. I: Interaction between MdPP2AC and MdSnRK2.6 measured by MST. Despite small changes in the mass of MdPP2AC, a signal-to-noise ratio of 29.30 enabled accurate determination of the interaction strength, and the affinity between MdPP2AC and MdSnRK2.6 was determined to be 109.09 ± 0.53 nM. Three independent experiments were performed with similar results.
Article Snippet: The blots were probed with a customized anti-MdPP2AC peptide-raised (
Techniques: Expressing, De-Phosphorylation Assay, Western Blot, Immunoprecipitation, Marker, Recombinant, Y2H Assay, Positive Control, Negative Control, FLC Assay, Bimolecular Fluorescence Complementation Assay, Pull Down Assay
Journal: Horticulture Research
Article Title: ALA reverses ABA-induced stomatal closure by modulating PP2AC and SnRK2.6 activity in apple leaves
doi: 10.1093/hr/uhad067
Figure Lengend Snippet: ALA treatment affects stomatal movement in transiently transgenic apple leaves . A : Stomatal aperture of apple leaves transiently overexpressing MdPP2AC or MdSnRK2.6 and treated with or without ALA. B : Stomatal aperture of apple leaves in which MdPP2AC or MdSnRK2.6 was transiently silenced, treated with or without ALA. Values are means ± SE of three biological replicates, and the same lowercase letters indicate no significant difference at p = 0.05.
Article Snippet: The blots were probed with a customized anti-MdPP2AC peptide-raised (
Techniques: Transgenic Assay
Journal: Horticulture Research
Article Title: ALA reverses ABA-induced stomatal closure by modulating PP2AC and SnRK2.6 activity in apple leaves
doi: 10.1093/hr/uhad067
Figure Lengend Snippet: Western blotting assay of phosphorylated MdSnRK2.6 and MdPP2AC protein abundances in OE- MdPP2AC apple leaves treated with exogenous ALA and/or ABA . A : Western blotting assay. B and C : Quantification of grey levels of the phosphorylated SnRK2.6 and PP2AC protein bands. Proteins were extracted from the apple leaf epidermis under different treatments before immunoprecipitation. Molecular mass (kDa) markers are shown on the left. All experiments were repeated at least three times with similar results. The same lowercase letters indicate no significant difference at p = 0.05
Article Snippet: The blots were probed with a customized anti-MdPP2AC peptide-raised (
Techniques: Western Blot, Immunoprecipitation