Fig 2: qRT-PCR analysis of gene expression patterns in Foxtail millet under drought stress. CK: control treatment. The SiActin gene was used as the internal control. The relative fold change abundance of corresponding proteins identified under drought stress compared with control treatment was presented in bottom line. NADP-ME: NADP-dependent malic enzyme, SiCAT: Catalase, SiLEA: late embryogenesis abundant protein, SiOEE: oxygen-evolving enhancer protein, GAPDH: glyceraldehyde-3-phosphate dehydrogenase, SiFBA: fructose-bisphosphate aldolase, QUINOR: quinone-oxidoreductase homolog, HSP70: heat shock protein, nsLTP: non-specific lipid-transfer protein, PPR: pentatricopeptide repeat-containing protein, Ribo: 60S ribosomal protein, SSADH: succinate-semialdehyde dehydrogenase, AOS: allene oxide synthase, PIP: aquaporin, ATPsyn: ATP synthase subunit O, PPDK: pyruvate, phosphate dikinase, Ampps: Xaa-Pro aminopeptidase, Tubu: tubulin, KCS: 3-ketoacyl-CoA synthase, PAP: plastid-lipid-associated protein, SiRLK: receptor-like serine/threonine-protein kinase

Fig 3: Cellular ATP contents and extrinsic proteasome-associated proteins are affected by oxidative and salt stresses. A, B Cellular levels of ATP, ADP (A) and ATP/ADP ratio (B) of PAG1-FLAG seedlings stress-treated for 24 h. Data are displayed as means ± SEM of six biological repeats. Significant differences between control and stress conditions were determined by independent Student’s t-tests (* = p < 0.05, ** = p < 0.01). C, D Co-immunoprecipitation (Co-IP) of PAG1-FLAG-tagged proteasomes from 12- and 24-h stress-treated seedlings via M2 anti-FLAG antibody resin. Cell lysis and co-IP were performed in the presence (C) and absence (D) of 20 mM ATP. Eluates are visualized on a silver-stained 12% SDS-PAGE gel. E Immunoblotting analysis with anti-HSP70, anti-FLAG, anti-RPN1, and anti-PA200 antibodies performed on Co-IP eluates from the 12-h time point shown in (C, D). For the immunoblots, Co-IP eluates were loaded alongside increasing concentrations of total lysate (TL) obtained from the 12-h control (C) sample (+ATP)

Fig 4: Co-immunoprecipitation assays of pF1β-1. The binding of RRL-pF1β-1 and RRL-1-77aaF1β-1 + mMORF3 to Hsp70 and 14-3-3 were strongly enhanced by LE treatment.

Fig 5: Analysis of Hsp70 content. (A) Electrophoresis and immunoblotting with anti-Hsp70 and anti-actin antibodies on proteins extracted from Giarraffa and Olivastra Seggianese plants, subjected to UV-B stress and collected at three selected time points (T0, T4, and T8). Lane 1: Olivastra Seggianese at T0. Lane 2: Giarraffa at T0. Lane 3: Olivastra Seggianese at T4. Lane 4: Giarraffa at T4. Lane 5: Olivastra Seggianese at T8. Lane 6: Giarraffa at T8. The same protein quantities were loaded in each lane. (B) Graph of the relative quantification of immunoblot intensities for Hsp70 relative to the actin content.