Journal: bioRxiv

Article Title: A novel repressor-activator-competitor module comprising C 2 H 2 zinc finger and NAC transcription factors regulates rice grain development

doi: 10.1101/2024.07.15.603577

Figure Lengend Snippet: (A) Schematic representation of ZOS1-15 and its truncated forms. ZF1 and ZF2 represent C 2 H 2 zinc fingers, DLN represents the repression motif, ΔN, ΔC and ΔDLN represent deletions of the respective regions. Diagram is not drawn to scale. (B) Y2H assays show homodimerization of ZOS1-15 and (C) its interacting partners, MED14_1, HDT702, OsTPL and ONAC024. _AD and _BD represent fusion with GAL4 activation domain or binding domain, respectively. SD/-LT indicates synthetic drop-out medium lacking leucine and tryptophan. SD/-AHLT indicates media lacking adenine, histidine, leucine, tryptophan and SD/-AHLT + X-α-gal indicates SD/-AHLT plus 5-bromo-4-chloro-3-indolyl-α-D-galactopyranoside (X-α-gal). p53_BD and T-antigen_AD are positive controls while Lam_BD and T-antigen_AD are negative controls. ND, 10 -1 , 10 -2 , 10 -3 denote no dilution, 10-fold dilution, 100-fold dilution and 1000-fold dilution respectively, of co-transformed yeast cultures. (D) BiFC assays show subcellular localisation of dimers of ZOS1-15 and its interactors in Nicotiana benthamiana leaf cells. YFP N and YFP C indicate N-terminal and C-terminal regions of YFP in fusion with the mentioned protein. Plasma membrane has been indicated by red arrow and nucleus is indicated by blue arrow. Left panel indicates reconstituted YFP signal at 514 nm. Merged indicate the merger of YFP and bright field and the right column shows the zoomed-in image of the interactions. Scale bar is 50 μm. Pull down and co-immunoprecipitation assays, respectively, confirm (E-F) homodimerisation of ZOS1-15 [GST_ZOS1-15 (44.92 kDa), YFP_ZOS1-15 (46.81 kDa)] and its interaction with (G-H) MED14_1 [GST_MED14_1 (74.51 kDa), YFP_ZOS1-15 (46.81 kDa)]; (I-J) HDT702 [MBP_HDT702 (66.26 kDa), GST_ZOS1-15 (44.92 kDa), YFP_ZOS1-15 (46.81 kDa)]; (K-L) TPL [MBP_OsTPL (74.07 kDa), GST_ZOS1-15 (44.92 kDa), YFP_ZOS1-15 (46.81 kDa)]; (M-N) ONAC024 [GST_ZOS1-15 (44.92 kDa), YFP_ONAC024 (62.54 kDa)]. Input, pull down fractions and the antibody used have been marked. Pull down and Co-IP lane with expected protein bands have been indicated by red arrow. IB denotes immunoblot.

Article Snippet: MBP-fused proteins were purified using MBP resin and the GST-tagged proteins were purified with glutathione agarose beads (G-biosciences ® ; USA).

Techniques: Zinc-Fingers, Activation Assay, Binding Assay, Transformation Assay, Membrane, Immunoprecipitation, Co-Immunoprecipitation Assay, Western Blot

Journal: bioRxiv

Article Title: A novel repressor-activator-competitor module comprising C 2 H 2 zinc finger and NAC transcription factors regulates rice grain development

doi: 10.1101/2024.07.15.603577

Figure Lengend Snippet: (A) Pull down assay showing interaction of MBP_ONAC024 (76.91 kDa) with GST_ONAC023 (53.97 kDa). (B) In-planta co-immunoprecipitation assay confirming the interaction of YFP_ONAC024 (62.54kDa) with ONAC023_MYC (39.97kDa). Pull down and Co-IP lanes with expected protein bands have been indicated by red arrows. IB denotes immunoblot. Graphs representing reproductive traits in 023_OE lines compared to WT; (C) average per grain length; (D) average per grain width and (E) average per grain weight (n = 50*3). (F) Transverse sections representing grain chalkiness in wild type (top), and 023_OE grains (5.1 – T3 grains; 36.1 – T4 grains); scale bar = 100µm. (G) Structural analysis of starch granules in WT and 023_OE lines by SEM, scale bar = 10µM; polyhedral starch granules have been marked in yellow and irregular shaped ones are in pink. (H) Estimation of AAC in mature seeds of WT and ONAC023 OE lines. Student’s t-test was used for calculating the significance ( *** = p-value< 0.005).

Article Snippet: MBP-fused proteins were purified using MBP resin and the GST-tagged proteins were purified with glutathione agarose beads (G-biosciences ® ; USA).

Techniques: Pull Down Assay, Co-Immunoprecipitation Assay, Western Blot, Starch

Journal: bioRxiv

Article Title: A novel repressor-activator-competitor module comprising C 2 H 2 zinc finger and NAC transcription factors regulates rice grain development

doi: 10.1101/2024.07.15.603577

Figure Lengend Snippet: Mature grains of 025_SOE (T3 grains) and WT arranged (A) lengthwise and (B) widthwise. Bar, 1 cm. Bar graphs show (C) average per grain length, (D) average per grain width and (E) yield per plant in WT and 025_SOE plants, (n= 50*3). (F) Transverse sections of WT and 025_SOE grains under the stereomicroscope. Scale bars, 1 mm. (G) Morphology profile of compound-type starch granules in the central endosperm sections of 025_SOE mature grains in comparison with WT. The transverse hand-cut sections of grains have been viewed under SEM. The polyhedral starch granules are outlined with yellow dashed lines and irregular granules have been marked in pink. Scale bars, 10µm. Line graph shows (H) AAC in WT and SS1/ ONAC025 transgenic grains and (I) TPC of WT and 025_SOE grains. The data represents two independent transgenic lines with three biological replicates each. The graphs depict average ± SE and statistical significance by one-tailed Student’s t-test ( * p<0.05, ** p<0.01, *** p<0.005). (J) Pull-down assay shows the interaction of MBP_SS1/ ONAC025 (75.8 kDa) with GST_MADS29 (54.86 kDa). The red arrow indicates the expected protein bands in the pull-down lane. IB, immunoblot. (K) Co-immunoprecipitation (Co-IP) assay confirming the interaction between SS1/ ONAC025 (61.43 kDa) and MADS29 (40.86 kDa). Input indicates total protein isolated from N. benthamiana leaves infiltrated with YFP_SS1/ ONAC025 and MADS29_MYC. The red arrow represents immunoprecipitated YFP_SS1/ ONAC025 in the protein mixture. IB denotes immunoblot.

Article Snippet: MBP-fused proteins were purified using MBP resin and the GST-tagged proteins were purified with glutathione agarose beads (G-biosciences ® ; USA).

Techniques: Starch, Comparison, Transgenic Assay, One-tailed Test, Pull Down Assay, Western Blot, Co-Immunoprecipitation Assay, Isolation, Immunoprecipitation

Journal: Journal of Functional Biomaterials

Article Title: Influence of the Application Time of Silane for the Bonding Performance between Feldspar or Lithium Disilicate Ceramics and Luting Resin Composites

doi: 10.3390/jfb14040231

Figure Lengend Snippet: Materials used in the present study.

Article Snippet: Luting resin composite , For MBP Specimen , Calibra translucent , Dentsply DeTrey GmbH Konstanz, Germany , 00041643 00023656.

Techniques: