Western blot analysis of selected candidate proteins (AtpB, 2Fe-2S, GLN2, Psb R, PRK, ALD) with their corresponding molecular weight in rice leaves during As stress under various S regimes (A); RuBisCo large subunit a band in coomassie blue staining (CBB) SDS gel served as a loading control (B), which used for normalization of detected proteins in densitometry studies (C) fold change indicated with respect of control samples.

Immunoblot analysis of photosynthetic complex accumulation in wild-type tobacco and the two ?psaI lines grown under low, intermediate, and high-light conditions. Because the accumulation of most tested proteins was highest under high-light conditions, lanes one to three contain samples diluted to 25%, 50%, and a 100% sample of wild-type tobacco grown under high-light conditions, to allow for semi-quantitative determination of changes in protein abundance. Lanes four and five contain the two transplastomic lines grown at 1000 µE m?2 s?1. Lanes six to eight contain wild-type tobacco and the mutants grown at intermediate light intensities, and lanes nine to eleven contain samples grown at low light intensities. For PSII, the accumulation of the essential subunits PsbB (CP43) and PsbD (D2) and the LHCB1 antenna protein were determined, while for the cytochrome b6f complex, the accumulation of the essential redox-active subunits PetA (cytochrome f), PetB (cytochrome b6), and PETC (Rieske FeS protein) was tested. AtpB was probed as an essential subunit of the chloroplast ATP. For PSI, in addition to the three essential plastome-encoded subunits PsaA, PsaB, and PsaC, the accumulation of the nuclear-encoded subunits PSAD, PSAH, PSAK, PSAL, and PSAN and of the four LHCI proteins (LHCA1, LHCA2, LHCA3, LHCA4) was determined. Finally, we examined the accumulation of Ycf4, the chloroplast-encoded PSI-biogenesis factor encoded in the same operon as PsaI, and the nuclear-encoded assembly factor Y3IP1.

Western blot analysis of PsbS and LHCSR proteins from dark-adapted (D, 30 min), light-treated (L, 2500 µmol photons m?2 s?1, 30 min) and post-dark-recovery chloroplasts (R, 1 h) isolated from B. corticulans. Samples were analysed by immunoblotting with an anti-PsbS (a), anti-LHCSR3 (b) and anti-LHCSR1 antibodies (c). Control samples are dark-adapted S. oleracea (a) and C. reinhardtii chloroplasts (b, c). Note that B. corticulans and S. oleracea samples have been probed against anti-PsbS antibody in the same membrane. The separation between lanes is only due to the presence of different samples between those presented of S. oleracea and B. corticulans (a). The ? subunit of ATP synthase (ATP-B) was used as loading control. 10 µg of Chl was loaded in each lane

LHCSR1 expression in isolated mutants following UV induction. (A) Gene expression analysis of LHCSR1 by RT-PCR. The CBLP gene was used as the housekeeping control. RNA and protein samples were collected after 1?hour of irradiation with UV. A 100-bp DNA ladder is shown. (B) Expression of the LHCSR1 protein was detected using specific antibodies, and the ATPB protein was used as the loading control. The protein samples were collected after 4?hours of irradiation with UV. Samples illustrated here are representative of three biological replicates.

CrCO and NF-YB are crucial for photoprotection. a The bleaching phenotypes of the reference strain (LHCSR1–Luc717) and the DSR mutants visualized in multiwell plates. Representative cell cultures treated with low light (LL; left wells) or high light (HL; right wells). Concentrations of the cultures were adjusted to 1.0?×? 107 cells/mL. b Chlorophyll content per cell after LL (closed bar) or HL (open bar) treatment of the cells shown in a. c Maximum quantum yield of photosystem II (Fv/Fm) during HL treatment. d qE quenching capability during HL treatment. e Immunoblot analysis of 3xFLAG-fused proteins (CrCO–FLAG and NF-YB–FLAG in crco-2/CrCO and nfyb-1/NFYB, respectively), LHCSR1, LHCSR3, and PSBS during HL treatment. ATPB protein levels are shown as the loading control. The experiments were performed three times with different biological samples (n?=?3 biological replicates; mean?± S.E.M); a representative experiment is shown in e

UVR8 interacts with COP1 and SPA1 to activate the CrCO-based transcriptional complex under UV irradiation. a Confocal images of UVR8–Venus–3xFLAG proteins in the DSR1–comp15 (uvr8/UVR8) strain. The cells were treated with UV for 30?min. Scale bars, 10?µm. b After 1?h of UV treatment, the cells were harvested and UVR8–Venus–3xFLAG proteins were immunoprecipitated by FLAG (M2) antibody with SURE-beads. The coimmunoprecipitated proteins were identified by LC-MS/MS analysis of the Coomassie Brilliant Blue-stained polypeptide bands obtained via SDS-PAGE separation after in-gel trypsin digestion. M, molecular mass standard. c Interaction profiles among COP1, SPA1, and CrCO visualized with Y2H assays, which were performed using COP1, SPA1, and CrCO fused with the AD and/or BD domains of GAL4. The culture conditions were as described for Fig. 2b. d Immunoblot analysis of LHCSRs, PSBS, and 3xFLAG-fused CrCO to visualize protein accumulation in the wild-type control (WT), crco-2, crco-2/CrCO, spa1, spa1 crco-2, and spa1 crco-2/CrCO strains before and after UV treatment. ATPB proteins were included as the loading controls

Immunoblot analysis of the photosynthetic proteins on the basis of equal total Ponceau S dye stained blot with proteins from leaves of wild type plants and pap1 mutant grown on MS medium in Petri dishes for four weeks under a 16 h light/8 h dark photoperiod at 23 °C with 100 ?E m?2 s?1. Proteins were visualized by immunoblotting using antibodies specific for RbcL, PsaB, PsbD, AtpB, RpoB, AccD and Lhcb2.4 proteins.

Western Blot analyses (a) and densitometric analyses of PsbS (b), VDE (c), PGRL1 (d), and cytf (e) proteins in leaves of tomato plants (Solanum lycopersicum L. cv. Malinowy Ozarowski) grown under different light conditions (see Material and Methods for details). The bands were normalized to the appropriate ? subunit of ATP synthase (ATPB) band (loading control, Figure S4) (a). The bar diagrams (b–e) represent pixel volumes (densitometric analyses) of proteins in samples. Each value represents the mean ± SD (n = 3) considering the control sample (WL) value as 1 (100%). Different letters indicate significant differences between treatments (p = 0.05) with a Tukey’s HSD test. MW—molecular weight.