Figures for Pituitary pathology and gene expression in acromegalic cats

Figure 1 Representative photomicrographs of growth hormone (A - C) and prolactin (D – F) immunostaining. A and D are x40 photomicrographs demonstrating specific immunostaining for somatrophs and lactotrophs, respectively. B and E are photomicrographs from a control cat C and F are from an acromegalic cat. <br> Figure 2 Representative images of SSTR2 immunoreactivity using feline pituitary tissue. A - D represent pituitary tissue exhibiting SSTR2 immunohistochemistry scores 0, 1, 2 and 3, respectively using the following criteria:.0 = absent; 1 = cytoplasmic staining; 2 = membranous staining in less than 50% cells or incomplete membranous staining; and 3 = circumferential membranous staining in &gt;50% cells. All presented photomicrographs collected at x100 magnification using Leica DM400 B, Leica Microsystems Cambridge, UK.<br> Figure 3 An electropherogram results from PCR products using multiplex 1 primer sets The blue peaks represent PCR products from gene specific primers and red peaks represent product size standards.<br> Figure 4 All images stained using Silver stain for reticulin fibres and counter stained using Nuclear Fast Red solution. A and C; reconstructed stitched pituitary x100 magnification photomicrographs from two control pituitaries. B and D; x400 magnification photomicrographs from A and C, respectively. The acinar pattern of reticulin staining is identified in B and D. This pattern of reticulin staining was demonstrated in all reticulin staining control pituitaries.<br> Figure 5 All images stained using Silver stain for reticulin fibres and counter stained using Nuclear Fast Red solution. A – D; selected images taken from reconstructed stitched pituitary x100 magnification photomicrographs from four HST pituitaries. A; disrupted reticulin staining and loss of acinar structure. B; areas of enlarged acini (blue stars) and areas of loss of acinar structure (blue cross). C; enlarged acini (blue stars) adjacent to normal sized and small acini. D; loss of acinar structure in the bottom right of the image (blue stars), and adenomatous tissue has compressed the normal pituitary tissue resulting in compression of the acini and a ring of cords of acini giving the impression of a pseudocapsule.<br> Figure 6 A: Bar charts comparing the relative gene expression of <i>SSTR1</i>, <i>SSTR2 and</i> <i>SSTR5</i> in pituitary tissue from control (CTRL) and acromegalic (Acro) cats determine using GeXP multiplex technique. <i>RPL18</i> is the reference gene. Bar height represents mean and error bars are 95% confidence intervals ** represents <i>P</i> &lt; 0.01 and *** represents <i>P</i> &lt; 0.001. Dot plot of the individual somatostatin profiles from each of the 19 acromegalic cats. <br> Table 1 Clinical data of cats in the control and acromegalic groups<br> Table 2 Gene expression data and GH, PRL and SSTR2 immunohistochemistry scoring of cats in the control and acromegalic groups.<br> Table 3 Summary of Spearman rank correlation gene expression data in the control group and acromegalic groups<br> Supplementary material 1Nucleotide sequences of primers used in multiplexes 1, 2 and 3.<br>Supplementary material 2Amino acid multiple sequence alignment for human and feline SSTR2<br>Supplementary material 3Amino acid multiple sequence alignment for porcine and feline growth hormone <br>Supplementary material 4Amino acid multiple sequence alignment for porcine and feline prolactin <br><br>