Histogram of the six measured traits in the F2:3 populations at Xunxian (a) and Zhengzhou (b) location.

<p>Histogram of the six measured traits in the F2:3 populations at Xunxian (a) and Zhengzhou (b) location.</p

Variance analysis of the six measured traits for shading and full light treatment in the F_2:3 populations at two locations.

<p>Note: L, location; B, block; G, genotype; S, shading treatment.</p>*<p>,**: the significant at the 0.05 and 0.01 levels.</p

The comparing value of the six measured traits for shading and full light treatment in the two parents, F₁ and F_2:3 populations at two locations.

<p>The comparing value of the six measured traits for shading and full light treatment in the two parents, F₁ and F_2:3 populations at two locations.</p

QTL detected for six measured traits related to shading under two treatments at two locations.

<p>Note: ^aAdditive effect; positive values of the additive effect indicate that the Zhong72 alleles are in the direction of increasing the traits. Dominance effect; positive values of the dominance effect indicate that the heterozygotes have higher phenotypic values than the respective means of the two homozygotes, and negative values indicate that heterozygotes have lower values than the means of the two homozygotes.</p>b<p>A, additive (d/a = 0.00–0.20); PD, partial dominance (d/a = 0.21–0.80); D, dominance (d/a = 0.81–1.20); OD, overdominance (d/a>1.20).</p>c<p>R² contribution rate.</p

The performance of the six measured traits for shade tolerance in the F_2:3 families under two treatments at two locations.

<p>Note:^aPH, plant height; EH, ear height; SD, stem diameter; DTT, day-to-tassel; DTS, day-to-silk; ASI, anthesis-silking interval.</p>b<p>The broad-sense heritability of stover yield and its nutrient components.</p>c<p>The confidence intervals of broad-sense heritability between 5% and 95% significant levels.</p

The four main climate factors in maize planting season and shading treatment season in 2008 at Xunxian (a) and Zhengzhou (b).

<p>The four main climate factors in maize planting season and shading treatment season in 2008 at Xunxian (a) and Zhengzhou (b).</p

Chromosomal location of quantitative trait loci (QTL) for shading sensitive related traits in maize under two shading treatments.

<p>The genetic distance in cM is listed on the left side of each chromosome.</p

Robust Immunity and Heterologous Protection against Influenza in Mice Elicited by a Novel Recombinant NP-M2e Fusion Protein Expressed in <em>E. coli</em>

<div><h3>Background</h3><p>The 23-amino acid extracellular domain of matrix 2 protein (M2e) and the internal nucleoprotein (NP) of influenza are highly conserved among viruses and thus are promising candidate antigens for the development of a universal influenza vaccine. Various M2e- or NP-based DNA or viral vector vaccines have been shown to have high immunogenicity; however, high cost, complicated immunization procedures, and vector-specific antibody responses have restricted their applications. Immunization with an NP–M2e fusion protein expressed in <em>Escherichia coli</em> may represent an alternative strategy for the development of a universal influenza vaccine.</p> <h3>Methodology/Principal Findings</h3><p>cDNA encoding M2e was fused to the 3′ end of NP cDNA from influenza virus A/Beijing/30/95 (H3N2). The fusion protein (NM2e) was expressed in E. coli and isolated with 90% purity. Mice were immunized with recombinant NM2e protein along with aluminum hydroxide gel and/or CpG as adjuvant. NM2e plus aluminum hydroxide gel almost completely protected the mice against a lethal (20 LD₅₀) challenge of heterologous influenza virus A/PR/8/34.</p> <h3>Conclusions/Significance</h3><p>The NM2e fusion protein expressed in <em>E. coli</em> was highly immunogenic in mice. Immunization with NM2e formulated with aluminum hydroxide gel protected mice against a lethal dose of a heterologous influenza virus. Vaccination with recombinant NM2e fusion protein is a promising strategy for the development of a universal influenza vaccine.</p> </div

Antibody response trend and long-term humoral immune response induced by NM2e protein in mice.

<p>(A and B) Mice were immunized intramuscularly with 10 µg of NM2e protein three times at 2-week intervals. Al(OH)₃ and/or CpG 1826 were used as adjuvants. Mice immunized with normal saline (NS) or adjuvant alone was used as negative controls. Serum was obtained from each mouse on days 14, 28, and 38, respectively, and analyzed for the presence of IgG antibodies specific for NP (left) or M2e (right), in an ELISA, as described in the Materials and Methods. Antibody response trends after three immunizations are presented in A, and the comparison of results on day 38 are presented in B. Columns show geometric mean antibody titers, and bars indicate the 95% confidence interval in each group. Plots in B show the NP- and M2e-specific IgG titers of all of the mice in each treatment group on day 38, and bars indicate the geometric mean antibody titers of each treatment group (<i>n</i> = 6 mice per experimental group, except <i>n</i> = 5 mice in the NS group). Lines above two or more groups indicate that they have the same comparative results. *, <i>p</i>≤0.05; **, <i>p</i>≤0.01; ***, <i>p</i>≤0.001 by one-way ANOVA. (C) Mice were immunized intramuscularly with 10 µg of NM2e protein formulated with Al(OH)₃ three times at 2-week intervals or immunized with a single dose of 10 µg of NM2e formulated with Al(OH)₃. Serum was prepared from each mouse at the indicated times, and NP- and M2e-specific IgG antibodies were analyzed by ELISA, as described in the Materials and Methods.</p

NM2e protein immunization schedule.

<p>The indicated mice were immunized intramuscularly with NM2e protein with or without adjuvant, three times at 2-week intervals. Blood was collected on days 14, 28, and 38, respectively. The immunized mice were challenged with influenza A virus PR8 at 20-fold the LD₅₀ on day 38. Body weight and survival were monitored for 3 weeks, until day 59.</p