Expression of NANOG and eGFP in mouse testes sections as detected by immunohistochemistry.

<p>(A–C) Expression of NANOG in mouse testis at different stages of spermatogenesis; Roman figures in seminiferous tubules represent the stage of the epithelial cycle; (D) Negative control section from which the primary antibody was omitted in the staining procedure; (E) Expression of eGFP in testis sections of a <i>Nanog</i> eGFP reporter; (F) eGFP staining on testis sections of a non-transgenic mouse, which served as a negative control for the staining in figures E. Abbreviations: AS =  type A spermatogonia, P =  Pachytene spermatocyte, T = Spermatid, ET =  elongating spermatids, S =  Sertoli cell, * =  non-specific binding to interstitial cells. Sections displayed in panels A–D were fixed in Bouins fixative; sections in panels E and F were fixed in formalin. Formalin fixation interfered with immunostaining of the seminiferous tubules.</p

Immunoblot analysis.

<p>Composite image of immunoblot results for NANOG on lysates of mouse ES cells and dog and pig testes. On the right is a Benchmark protein ladder (Invitrogen) and the indicated molecular weights for each band. Abbreviations: mES =  mouse embryonic stem cells, <i>Cf</i> =  <i>Canis familiaris</i>, <i>Ss</i> =  <i>Sus scrofa</i>.</p

Antibodies used for immuno applications.

<p>AA =  Amino Acids, IHC =  immunohistochemistry, IF =  immunofluorescence, IB = immunoblotting.</p

Expression of <i>Nanog</i> in testis of various mammalian species as determined by RT-PCR.

<p>(A) Expression of <i>Nanog</i> in mouse testis of the Black 6 strain (BL/6) and the SV129 strain (SV129) (B) Expression of <i>eGFP</i> in testis of a TNG mouse; (C) Expression of <i>NANOG</i> in dog and pig testes; (D) Expression of <i>NANOG</i> in human testis; in all RT-PCR experiments, amplicons were of the expected sizes (See also <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0010987#pone-0010987-t002" target="_blank">Table 2</a>) and products were identified by sequence analysis. Abbreviations: TNG =  testis cDNA from <i>Nanog</i> eGFP reporter mouse; <i>Cf</i> =  <i>Canis familiaris</i>, <i>Ss</i> =  <i>Sus scrofa</i>, <i>Hs</i> =  <i>Homo sapiens</i>; +  = cDNA synthesis reaction performed with reverse transcriptase, −  =  minus reverse transcriptase control.</p

Expression of NANOG in human testis as determined by immunofluorescence in paraffin-embedded sections.

<p>(A–C) NANOG expression in differentiating male germ cells; dashed lines mark the boundaries of tubules.</p

Expression of NANOG in pig testis as determined by immunofluorescence.

<p>(A–C) NANOG expression in differentiating male germ cells; (D–F) high magnification image of NANOG expression in pachytene spermatocytes; (G–I) image of section in which the primary antibody was replaced by blocking solution, to control for non-specific binding of the secondary antibody; (J–K) representative high magnification image of testis section that was incubated with a rabbit IgG isotype control, to control for non-specific binding of the primary antibody. Dashed lines in panels A–C and G–I mark the boundaries of the tubules.</p

Primers used for RT-PCR and sequence analysis.

<p>F =  forward primer, R =  reverse primer. Ta =  annealing temperature used in PCR reaction.</p

Education for sustainable development: A study in adolescent perception changes towards sustainability following a strategic planning-based intervention: The young persons’ plan for the planet program

In 2016, the United Nations (UN) launched the 17 Sustainable Development Goals (SDGs) as a framework for sustainable development and a sustainable future. However, the global challenge has been to engage, connect, and empower communities, particularly young people, to both understand and deliver the 17 SDGs. In this study, we show the benefit of a strategic planning-based experiential learning tool, the Young Persons’ Plan for the Planet (YPPP) Program, to improve the underlying competencies of Australian and Mauritian adolescents in increasing understanding and delivering the SDGs. The study was conducted with 300 middle to senior high school students, in 25 schools throughout Australia and Mauritius, over an 18-month period. The intervention included the development of research, strategic planning, management, STEM (Science Technology, Engineering, Maths) and global competency skills in the students, to enable them to build and deliver regional and national SDG plans. Research methods included pre- and post-intervention testing of the attitudes of these students to sustainable development outcomes and compared these attitudes to subsets of scientists and the Australian national population. Our results, from both qualitative and quantitative evidence, demonstrate significant improvements in these adolescents’ appreciation of, and attitudes towards, the SDGs and sustainable outcomes, across a range of key parameters. The results from the 76 students who attended the International Conference in Mauritius in December 2018 demonstrate significant improvements in mean levels of understanding, and attitudes of the students towards the SDGs awareness (+85%), understanding/engagement (+75%), motivation (+57%), and action orientation/empowerment (+66%). These changes were tested across a range of socio-demographic, geographic, and cultural parameters, with consistent results. These findings have significant implications for the challenge of sustainable education and achieving community engagement and action towards the SDGs in Australia and Mauritius, particularly for young people. As the intervention can be replicated and scaled, the findings also highlight the opportunity to extend both the research and this type of experiential learning intervention across both broader geographies and other generation and community segments