Figure 6: E. carinatus venom does not induce local tissue damage or NETosis in neutropenic mice.

(a) Representative photographs of mice 8 h after E. carinatus venom (LD₅₀) injection in the tail show intense venom-induced wound in the tail of a normal mouse (left), but no haemorrhage in a neutropenic mouse (right). Scale bars, 2 cm. (n=10). (b) The corresponding tail injury score 8 h after the E. carinatus venom (LD₅₀) injection is shown in a bar graph. The data are presented as mean±s.e.m. n=6 venom-injected normal mice; n=10 venom-injected neutropenic mice. **P<0.01; Student’s t-test. (c) Western blot analysis of the appearance of H3Cit (top) in tail tissue homogenates taken from normal and neutropenic mice 8 h after E. carinatus venom (LD₅₀) injection. The quantification of H3Cit levels compared with H3 levels is shown (bottom). AU, arbitrary units; H3Cit, citrullinated histone 3; H3, histone 3. The data are presented as mean±s.e.m. (n=4). ***P<0.001 versus the normal control mice; one-way analysis of variance, followed by Dunnett’s post-hoc test. The PVDF membranes were cut based on molecular weight of respective protein using protein molecular weight marker and then probed with respective antibodies. (d) H&E-stained tail tissue sections from neutropenic (top right) and normal mice (bottom left) injected with E. carinatus venom (LD₅₀); the yellow portion is enlarged and shown on the right. Tissue from PBS-injected normal mice is also shown (top, left). Scale bars, 100 μm. (n=4). (e) Representative immunofluorescence images of neutropenic mouse tail tissue 8 h after E. carinatus venom (LD₅₀) injection, focused beneath the epithelial layer. Scale bars, 100 μm (n=4).