Growth model parameter estimates.

<p>Parameter estimates (with 95% CI) for the sex effect on growth from the best model (i.e., lowest AIC_c) for each morphological measurement of Adélie Penguin chicks during 2012–13 and 2013–14 on Ross Island, Antarctica. Plus signs denote additive effects and asterisks denote interactions. Sex was not supported as an important factor associated with foot growth.</p

Estimated growth rates.

<p>Average daily growth rate estimates (with 95% CI) from best model (i.e., model with lowest AIC_c) relating growth rates of morphological characteristics and mass to sex, year, brood size, chick hatching order; and sample sizes (n) for male and female Adélie Penguin chicks measured, and weighed during 2012–13 (2012) and 2013–14 (2013) on Ross Island, Antarctica. Sex was not supported as an important variable for foot growth so only means by year (best model results) are reported.</p

Measurement diagram.

<p>Schematic illustrating the location of morphological measurements collected from male and female Adélie Penguin chicks during Austral summers of 2012–13 and 2013–14 on Ross Island, Antarctica. Bill was measured to the nearest hundredth mm and remaining measurements to the nearest mm. Dashed line indicates humeral head on underside of the flipper.</p

Characterization of kinesin switch I mutations that cause hereditary spastic paraplegia

<div><p>Kif5A is a neuronally-enriched isoform of the Kinesin-1 family of cellular transport motors. 23 separate mutations in the motor domain of Kif5A have been identified in patients with the complicated form of hereditary spastic paraplegia (HSP). We performed in vitro assays on dimeric recombinant Kif5A with HSP-causing mutations in the Switch I domain, which participates in the coordination and hydrolysis of ATP by kinesin. We observed a variety of significantly reduced catalytic and mechanical activities as a result of each mutation, with the shared phenotype from each that motility was significantly reduced. Substitution of Mn²⁺ for Mg²⁺ in our reaction buffers provides a dose-dependent rescue in both the catalytic and ensemble mechanical properties of the S203C mutant. This work provides mechanistic insight into the cause of HSP in patients with these mutations and points to future experiments to further dissect the root cause of this disease.</p></div

Effect of divalent cation on S203C microtubule gliding velocity.

<p>A) Microtubule gliding velocity of the S203C mutant in buffers with the divalent cations shown. The asterisk indicates that the velocity in Mn²⁺ is significantly greater than the velocity in Mg²⁺ (p<0.01), although still significantly lower than wild type in Mg²⁺ (p<0.01). Note the log scale of the Y-axis. B) Microtubule gliding velocity of the S203C mutant that was purified in the presence of Mg²⁺ (Mg-S203C) and Mn²⁺ (Mn-S203C). The single asterisk indicates that the velocity of Mg-S203C in Mn²⁺ is significantly greater than in Mg²⁺ (p<0.01), while the double asterisk indicates that the velocity of Mn-S203C in Mn²⁺ is significantly greater than in Mg²⁺ (p<0.01), and also significantly greater than the velocity of Mg-S203C in Mn²⁺ (p<0.05). C) Histograms of the data for S203C mutants in the conditions indicated. A similar number of microtubules (n = 90–110) were analyzed in each condition.</p

Molecular dynamics simulations of predicted altered kinesin structure.

<p>Images shown are the end point of a 10 ns simulation for wild type or mutant kinesin using the 4HNA structure as a starting point. Amino acids of interest are labeled in white. Intramolecular distances between potential bonding partners are shown in purple, with units of angstroms.</p

Microtubule gliding velocity.

<p>Microtubule gliding velocity.</p

Effects of HSP mutations on the basal and MT-Stimulated ATPase rate.

<p>Effects of HSP mutations on the basal and MT-Stimulated ATPase rate.</p

MT affinity in the presence of AMPPNP.

<p>MT pelleting assays were performed as described for <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0180353#pone.0180353.g001" target="_blank">Fig 1</a>, except 5 mM AMPPNP was substituted in the reaction buffer. Experiments were repeated in a range of tubulin concentrations from 0 to 20 μM and the results of the densitometry analysis of the western blots produced are plotted for each Kif5A mutant. The curves shown are the best fit to the data using the exponential decay function in Origin 8 in a user free computational method. A) Curves of fraction Kif5A bound as a function of tubulin concentration for the wild type and mutant proteins. B-H) Individual Kif5A curves (WT or mutant as indicated) with mean +/- SD for each concentration of tubulin analyzed.</p

The Anoikis Effector Bit1 Displays Tumor Suppressive Function in Lung Cancer Cells

<div><p>The mitochondrial Bit1 (Bcl-2 inhibitor of transcription 1) protein is a part of an apoptotic pathway that is uniquely regulated by integrin-mediated attachment. As an anoikis effector, Bit1 is released into the cytoplasm following loss of cell attachment and induces a caspase-independent form of apoptosis. Considering that anoikis resistance is a critical determinant of transformation, we hypothesized that cancer cells may circumvent the Bit1 apoptotic pathway to attain anchorage-independence and tumorigenic potential. Here, we provide the first evidence of the tumor suppressive effect of Bit1 through a mechanism involving anoikis induction in human lung adenocarcinoma derived A549 cells. Restitution of Bit1 in anoikis resistant A549 cells is sufficient to induce detachment induced-apoptosis despite defect in caspase activation and impairs their anchorage-independent growth. Conversely, stable downregulation of Bit1 in these cells significantly enhances their anoikis resistance and anchorage-independent growth. The Bit1 knockdown cells exhibit significantly enhanced tumorigenecity <i>in vivo</i>. It has been previously shown that the nuclear TLE1 corepressor is a putative oncogene in lung cancer, and we show here that TLE1 blocks Bit1 mediated anoikis in part by sequestering the pro-apoptotic partner of Bit1, the Amino-terminal Enhancer of Split (AES) protein, in the nucleus. Taken together, these findings suggest a tumor suppressive role of the caspase-independent anoikis effector Bit1 in lung cancer. Consistent with its role as a tumor suppressor, we have found that Bit1 is downregulated in human non-small cell lung cancer (NSCLC) tissues.</p></div