PMC

(A) Comparison of SEC patterns of tetraPEGylated canine Hb (the major component eluting from the Q-Sepharose HP column) with that of HbA, and hexaPEGylated Hb generated using thiolation-mediated maleimide chemistry-based PEGylation maleimide PEG5K []. (B) RP-HPLC pattern of canine Hb. (Upper curve) PEGylated canine Hb before chromatographic purification. (Lower curve) PEGylated canine Hb after chromatographic purification on Q-Sepharose HP. (C) SDS gel patterns of tetraPEGylated canine Hb before and after Q-Sepharose chromatography and (SP-PEG5K)₂-Hb. The tetraPEGylated canine Hb carries one major band. The mobility of this band corresponds to that of human β-globin PEGylated with PEG5K at its Cys⁹³(β), present in the sample of diPEGylated Hb. Lane 1, protein markers; lane 2, canine Hb; lane 3, tetraPEGylated canine Hb before Q-Sepharose purification; lane 4, tetraPEGylated Hb after purification on the Q-Sepharose column; and lane 5, diPEGylated human Hb. (D) IEF pattern of PEGylated canine Hb. Lane 1, canine Hb; lane 2, tetraPEGylated canine Hb after purification on a Q-Sepharose column; lane 3, diPEGylated human Hb; lane 4, hexaPEGylated Hb generated by EAF PEGylation; lane 5, control human Hb.

A) Expression of latexin in non-stimulated canine M0-macrophages. B) Expression of latexin in canine M1-macrophages. C) Expression of latexin in canine M2-macrophages. D) Expression of MS4A2 in canine M0-macrophages. E) Expression of MS4A2 in canine M1-macrophages. F) Expression of MS4A2 in canine M2-macrophages. A-F) Scale bars = 100 μm. Nuclear counterstaining with bisbenzimide. G) Dot plots illustrating a significantly higher percentage of immunopositive cells in canine M1- and M2-macrophages as compared to M0-macrophages (One-factorial ANOVA with group-wise t-tests, astrerisk = p<0.05). H) Dot plot showing lack of statistical differences in the percentage of immunopositive cells for MS4A2 in canine M0-, M1-, and M2-macrophages.

The upper canine morphology and replacement process of sabertooth taxa (above) versus the upper canine morphology and replacement process of closely related conical‐toothed taxa (below). The upper canine replacement process of three sabertooth lineages involves permanent upper canine (C¹) eruption medial, within the lingual concavity of the deciduous upper canine (dC¹). The upper canine replacement process of related conical‐toothed taxa consists of the permanent upper canine (C¹) erupting anterior and medial to the deciduous upper canine (dC¹). (a) Nimravid, AMNH FM 62013. Natural fractures of the upper canine teeth at the alveolar border showing cross sections of the right permanent upper canine (C¹) erupting within the lingual concavity of the right deciduous upper canine (dC¹), ventral view. (b) Barbourofelid, AMNH FM 61895 (Barbourofelis morrisi). Damaged maxilla and natural fractures of the upper canine teeth revealing the right permanent upper canine (C¹) developing lingual to the right deciduous upper canine (dC¹), anteroventral view. (c) Felid, UCMP 152566 (Smilodon fatalis). Micro‐CT slice of the upper canine teeth showing cross sections of the right permanent upper canine (C¹) within the lingual concavity of the right deciduous upper canine (dC¹), ventral view. Modified from Micro‐CT data Wysocki, Feranec, Tseng, and Bjornsson (). (d) Mustelid, USNM 222659 (Taxidea taxus). Right permanent upper canine (C¹) erupting anterior and medial to the right deciduous upper canine (dC¹), ventral view. Modified from Long (). (e) Felid, AMNH 17420 (Panthera tigris). Left permanent upper canine (C¹) erupting anterior and medial to the left deciduous upper canine (dC¹), ventral view. The natural fracture of the deciduous upper canine (dC¹) demonstrates the conical morphology that lacks the marked lingual concavity and mediolateral compression found in the deciduous upper canines of the sabertooth taxa. Scale bars = 1 cm

(A-D) Photomicrographs of sections of canine embryos at 22, 25, 27–28 and 30 days post-fertilization (dpf). (A1-D1) Histological section of male canine gonadal ridge during the early gestation period (22–30 dpf) (Scale bars are 50 μm). Sections of putative PGCs at 22, 25, 27–28 and 30 dpf immunoassayed for the early germ cell marker (nuclear) POU5F1 (red) and the late germ cell marker (cytoplasmic) DAZL (green) (Scale bars are 50 μm). (E) Percentage of quantified canine putative PGCs in canine gonads between 22 and 50 dpf detected using the POU5F1 and DDX4 antibodies. (F) Percentage of quantified canine putative PGCs in canine gonads between 22 and 50 dpf detected using the POU5F1 and DAZL antibodies. (G) Percentage of quantified canine putative PGCs in canine gonads between 22 and 50 dpf detected using the POU5F1 and DPPA3 antibodies.