Journal: Frontiers in Cellular Neuroscience

Article Title: The prion protein constitutively controls neuronal store-operated Ca 2+ entry through Fyn kinase

doi: 10.3389/fncel.2015.00416

Figure Lengend Snippet: The Tyr-kinase Fyn is the link between PrP C and SOCE. (A) PrP C reduces the level of active Fyn, as evident from both the representative WB (upper panel) of PrP-Tg and PrP-KO CGN probed with an antibody to p-SFK or total Fyn (both run in duplicate), under SOCE-activating conditions (Ca 2+ -depleted stores), and the corresponding densitometric analysis of p-SFK immunosignals normalized to that of total Fyn (lower panel). Contrary to other neuronal SFK members (Supplementary Figure 4), the identical apparent mass of p-SFK and Fyn indicates that the Fyn band corresponds to the p-SFK band. Similar results were obtained under basal conditions, i.e., with Ca 2+ -filled stores (see Supplementary Figure 3). (B,C) Addition (+) of the SFK inhibitors saracatinib (sara, 5 μM) and PP2 (10 μM), but not of PP3 (10 μM), reduces the level of both p-SFK (B) and total Tyr-phosphorylated (p-Tyr) proteins (C) of CGN compared to the untreated (−) samples. (B) The upper panel reports a representative WB of the two neuronal genotypes treated with, or without, the SFK inhibitors, and immunostained as in (A) , while the lower panel shows the corresponding densitometric analysis of p-SFK normalized to the band intensity of total Fyn. Veh (vehicle) indicates the control experiment run in the presence of DMSO (0.1%). (C) The upper panel reports the WB of total p-Tyr proteins present in the two CGN genotypes treated as in (B) . In the corresponding densitometric analysis (lowest panel), the p-Tyr band intensity was normalized to that of the Coomassie blue-stained bands (middle panel). (D) Only saracatinib and PP2 decrease SOCE-induced [Ca 2+ ] pm peaks, and abrogate the difference observed in control (veh-treated) PrP-Tg and PrP-KO CGN. (E) STIM1 is more abundantly Tyr-phosphorylated in PrP-KO CGN than in PrP-Tg CGN under SOCE-stimulating conditions. This is evident from the representative WB (left panel) of immunoprecipitated (IP) STIM1 probed with an antibody to either p-Tyr or total STIM1 (arrow), and from the corresponding densitometric analysis (right panel) reported as the ratio between the p-Tyr band intensity and the STIM1 band intensity. The arrowhead in the left panels indicates the immunosignal of the mouse IgG used in the immunoprecipitation assay. On the left of the WB, MW standards are indicated. * p < 0.05, ** p < 0.01, *** p < 0.001, Student’s t -test. The analysis of the statistical significance ( p -value, Student’s t -test) of data for the comparison between different treatments within each PrP genotype (B–D) is reported in Supplementary Table 2. Other details are as in the legend to Figure .

Article Snippet: Instead, the SFK inhibitors [PP2 (10 μM, Tocris) and saracatinib (5 μM, Santa Cruz Biotechnology)], and the negative control of PP2, PP3 (10 μM, Santa Cruz Biotechnology), were added during the reconstitution step, and kept in the perfusion buffer throughout the entire experiment.

Techniques: Control, Staining, Immunoprecipitation, Comparison

Journal: Frontiers in Plant Science

Article Title: Final Selection of Quality Protein Popcorn Hybrids

doi: 10.3389/fpls.2021.658456

Figure Lengend Snippet: Description of cultivars tested in 2020 summer trials.

Article Snippet: These four QPP hybrids were also derived from the same PP1 x PP3 (H13) ConAgra pedigree, which did not have correspondingly lower popping quality trait values compared to the other ConAgra varieties ( ).

Techniques:

Journal: Frontiers in Plant Science

Article Title: Final Selection of Quality Protein Popcorn Hybrids

doi: 10.3389/fpls.2021.658456

Figure Lengend Snippet: Protein profiling of QPP and ConAgra elite lines. (A) Principle component analysis (PCA) of protein-bound amino acids in raw flour of ConAgra elite lines, B73 for reference, and QPP hybrids revealed a distinct segregation between QPP and original popcorn-derived cultivars. B73 grouped with ConAgra lines H12, H13, and H14, while H11 and H15 independently segregated with QPP H2. QPP H2 had a distinct proteome compared to all other QPP hybrids and similar lysine levels compared to ConAgra lines. (B) Protein-bound lysine in raw flour of all genotypes revealed significantly higher lysine levels in all QPP lines except H2. (C) Zein extraction and SDS-PAGE analysis of randomly selected kernels revealed a significant reduction in 22-kD-alpha zein, varying production of 19-kD-alpha zein, and increased expression of the 27-kD γ-zein in QPP lines compared to ConAgra lines, consistent with a homozygous opaque-2 profile. Compositely, these results verified the successful introgression and stabilization of the homozygous mutation in the BC 2 F 5 and BC 3 F 4 QPP populations.

Article Snippet: These four QPP hybrids were also derived from the same PP1 x PP3 (H13) ConAgra pedigree, which did not have correspondingly lower popping quality trait values compared to the other ConAgra varieties ( ).

Techniques: Derivative Assay, Extraction, SDS Page, Expressing, Mutagenesis

Journal: Frontiers in Plant Science

Article Title: Final Selection of Quality Protein Popcorn Hybrids

doi: 10.3389/fpls.2021.658456

Figure Lengend Snippet: Select trait measurements of cultivars tested in 2020 summer trials.

Article Snippet: These four QPP hybrids were also derived from the same PP1 x PP3 (H13) ConAgra pedigree, which did not have correspondingly lower popping quality trait values compared to the other ConAgra varieties ( ).

Techniques:

Journal: Frontiers in Plant Science

Article Title: Final Selection of Quality Protein Popcorn Hybrids

doi: 10.3389/fpls.2021.658456

Figure Lengend Snippet: 2020 Ranking System Selection Index Results. Utilization of the 2020 Ranking System enabled a visual display of overall cultivar ranking from best to worst, left to right, respectively. Color by variable identified individual hybrid pitfalls (the longer the stacked column, the farther from the best hybrid) and high trait values. H13, PP1 x PP3, ranked highest out of all hybrids, scoring relatively lower only because of its lack of protein-bound lysine content. QPP H10 ranked the best compared to all other QPP lines and ranked higher than H14, or Commercial Line 1. QPP BC 2 -derived hybrids H3, H4, and H1 ranked, respectively, higher than the rest, while BC 3 -derived hybrids H6, H9, and H7 all ranked higher than original popcorn hybrid line H12. H8, H5, and H2 ranked lowest out of all hybrids. Economic weights for each trait were determined to reflect consumer and producer interests in a ready-to-eat Quality Protein Popcorn product. Lysine levels, yield, and expansion volume were considered equally important traits while ranking all hybrids.

Article Snippet: These four QPP hybrids were also derived from the same PP1 x PP3 (H13) ConAgra pedigree, which did not have correspondingly lower popping quality trait values compared to the other ConAgra varieties ( ).

Techniques: Selection, Derivative Assay