Haemoglobin measurements.

<p>Haemoglobin (Hb) is measured in the blood from the mice. Tg mice (open symbols) are anaemic compared to Wt mice (solid symbols), but the haemoglobin levels are restored in Tg mice 2 weeks after injection.</p

Epo expression.

<p><b>A:</b> Epo protein is significant higher in serum samples after 11 weeks for both Tg (<i>n =</i> 10) and Wt (<i>n =</i> 6) mice receiving Epo 1.25 μg/mouse compared to their Controls (<i>n =</i> 11 and <i>n =</i> 9, respectively) (^<i>#</i><i>P</i> = 0.004 and <i>*P</i> < 0.001, ± SD). <b>B:</b> Epo mRNA expression from kidneys. Significant decreased expression is seen in Tg Control (<i>n =</i> 13) compared to Wt Control (<i>n =</i> 11) (^<i>¤</i><i>P</i> = 0.025, ± SD). Treatment with Epo plasmid decreases endogenous Epo expression in both Tg and Wt mice (^{<i>§</i>,<i>†</i>}<i>P</i> < 0.001, ± SD).</p

Evaluation of plasmid expression site and level.

<p><b>A:</b> Plasmid signal day 8 after injection in Tg Control (<i>n =</i> 8) and Wt Control (<i>n =</i> 5) corresponding to the location of the liver. Scanning Control mice without preceding injection with luciferin are scanned simultaneously (<i>n =</i> 4). <b>B:</b> Regions of interest marks the signal and is measured as total flux (photons/second) (± SD). There is a tendency to higher plasmid expression in Wt Control mice (<i>n =</i> 5) than Tg Control mice (<i>n =</i> 8) (<i>P</i> = 0.13). <b>C:</b> Expression of Epo mRNAfrom the liver at termination of the experiment. Both Tg (<i>n =</i> 11) and Wt (<i>n =</i> 11) mice treated with Epo express significant more plasmid from the liver compared to their Controls (^{<i>¤</i>,<i>§</i>}<i>P</i> < 0.001, ± SD). Significant difference between Tg Epo 1.25 μg/mouse (<i>n =</i> 11) and Wt Epo 1.25 μg/mouse (<i>n =</i> 11) (^<i>#</i><i>P</i> < 0.001, ± SD) are found.</p

Expression and content of fibronectin in kidneys.

<p><b>A:</b> Transgenic Control mice (<i>n =</i> 13) express more fibronectin mRNA in total kidney tissue compared with Wt Control mice (<i>n =</i> 11) (<i>*P</i> < 0.001, ± SD), but treatment with Epo has no effect in either type (<i>n =</i> 11 for both types). <b>B:</b> Total fibronectin protein is present in increased amounts in transgenic mice (^<i>#</i><i>P</i> < 0.001, ± SD), but treatment with Epo has no effect. A representative section of the western blot is shown.</p

Evaluation of the tubular basement membrane (TBM).

<p><b>A:</b> Thickening of TBM evaluated by electron microscopy. Arrows indicate TBM thickness. <b>B:</b> Collagen α1/α2(IV) mRNA expression is increased in Tg Control (<i>n =</i> 8) compared to Wt Control mice (<i>n =</i> 5) (<i>*P</i> < 0.001, ± SD), but treatment with Epo has no effect in either type. C: Collagen αI(III) mRNA expression is increased in Tg (<i>n =</i> 8) compared to Wt mice (<i>n =</i> 5) (^<i>§</i><i>P</i> < 0.01, ± SD), but treatment with Epo has no effect in either type.</p

The gecko tail is segmental in nature. A)

<p>A histochemical analysis of a longitudinal <i>Gekko Gecko</i> tail section is shown (Mallory’s trichrome stain). The connective tissues and scales appear blue, the muscles appear red, and the adipose tissues appear white. The data illustrate the segmented structure of the gecko tail. The inserts focus on the areas where the collagen fibers are condensed. <b>B)</b> A histochemical analysis focusing on the proposed autotomy septum in the dermis. The arrows show the position of the septum in the condensed collagen area. In addition, arrows indicate the fracture plane from the condensed collagen area to the indentation between two scales where the severing takes place. <b>C)</b> Electron microscopy analysis focusing on the proposed autotomy septum in the condensed collagen area. The analysis reveals the presence of embedded cells in the proposed fracture plane that may function as a “cellular zipper”.</p

Wedge-shaped extensions project from the proximal end of the released tail stump. A)

<p>The proximal part of a released tail stump is observed from the side. The picture shows the wedge-shaped extensions. <b>B)</b> A wedge-shaped extension. <b>C)</b> The distal part of the tail that remains on the animal. The grooves, in which the extensions are fitted, in an end-to-end tapered “finger joint” fashion, are only vaguely seen just below the dermis. Immediately after autotomy, the skin contracts around the end of the tail stub, making the grooves difficult to observe.</p

Cross-fracture-plane structures are absent in the gecko tail.

<p>Magnetic resonance imaging was used to analyse a detached tail stump (see also Movies S1–S3). The data revealed the structures of the muscle fibers in the different segments of the gecko tail. Two areas have been encircled to point out the position of the muscle fiber termini neighbouring the autotomy fracture plane. The image illustrates the expanded nature of these termini. The presence of these “mushroom-shaped” structures in the fracture plane and the absence of these structures in the interior muscle fibers illustrate the conformational change of the muscle termini that occurs during autotomy. The interdigitating arrangement of the muscle fibers are observable and the picture also shows that no through-going structures between muscle segments are observable, thereby supporting the concept that tail segments interact by adhesion forces.</p

The muscle fibres terminate in “mushroom-shaped” structures.

<p>Scanning electron microscopy analyses of the wedge-shaped extensions (cranial margins of individual tail muscles) projecting from the proximal end of the released tail stump demonstrated the presence of “mushroom-shaped” structures at the termini of the muscle fibres after autotomy. These structures are present on all sides of the extensions except on the outer part.</p

Relative gene expression of ZnT-3 and ZnT-8 after 24 hours of 100 µM DEDTC treatment.

<p>INS-1E cells were treated with DEDTC at 5 mM glucose. A) ZnT-3 gene expression normalised to Cltc, HPRT and HSPcb. Data are mean and SEM (*p<0.05). N = 6. B) ZnT-8 gene expression normalised to Cltc, HPRT and HSPcb. Data are mean and SEM (*p<0.01). N = 6.</p