Journal: Inflammation and Regeneration

Article Title: Resolving the conflicts around Par2 opposing roles in regeneration by comparing immune-mediated and toxic-induced injuries

doi: 10.1186/s41232-022-00238-2

Figure Lengend Snippet: Par2 expression is maintained in WT hepatocytes and not in WT leukocytes 14 days after ConA injection. The core of large infiltrates is composed of apoptotic cells, which are CD45, Par2, and albumin negative. The apoptosis is indicated by the loss of the normal nuclei structure (note the ring-like circle of the Dapi staining, most apparent in C . The outer layer of the mononuclear infiltrate is composed of CD45+ leukocytes (shown in red). At day 14, these cells do not express Par2 and present normal nucleus. Healthy hepatocytes are not present in the infiltrate and observed by the uneven albumin staining of the infiltrate. At this stage of the ConA model, all apparent Par2+ cells express albumin (most observed in A and C ’). The albumin+ staining is an indication of a functional hepatocyte. A Low, B middle, and C high magnifications. Scale bar = 50μm

Article Snippet: Nuclei were visualized with DAPI Fluoromount-G TM (Bar Naor Ltd, Ramat Gan, Israel).

Techniques: Expressing, Injection, Staining, Functional Assay

Journal: PLOS Pathogens

Article Title: A divergent cyclic nucleotide binding protein promotes Plasmodium ookinete infection of the mosquito

doi: 10.1371/journal.ppat.1013467

Figure Lengend Snippet: (A) PbΔCBP-O ookinetes show regular motility. Average speed is 3.07 and 3.2 μm/min for wild-type and ΔCBP-O, respectively (wild-type n = 38; PbΔCBP-O n = 48, representative data from 3 independent experiments, unpaired t-test). (B) Comparable ookinete maturation in the mosquito midgut is observed in PbΔCBP-O and wildtype parasites. Nineteen or twenty-one hours post-infection, six midguts were scored per replicate (n = 2). Significance was analysed using an unpaired t-test. (C) Expression of micronemal proteins including CTRP, WARP and chitinase are unaffected in the absence of CBP-O. Actin is used as loading control (left panel). Chitinase secretion is maintained in PbΔCBP-O ookinetes. Western blot of culture supernatants from wild type and PbΔCBP-O ookinetes show similar levels of chitinase secretion (right panel), representative data from 2 independent experiments, (D) Immunofluorescence showing decreased ookinete presence in the basal side of mosquito midguts. Midguts were fixed and stained with P25 (plasma membrane marker) and DAPI 24-25 hours post-infection. P25 + parasites were quantified, and single dots show number of ookinetes counted per midgut (n = 20 midguts per genotype, unpaired t-test.). Scale bar = 200μm. (E) Quantification of salivary gland sporozoites. Each data point is an average of five infected mosquitoes (representative data from two independent experiments). (F) Expression of An. stephensi SRPN6 mRNA relative to An. stephensi rps7 in mosquitoes that were fed uninfected blood (control), fed with wt or PbΔCBP-O infected blood (Data are mean ± SD, n = 25 midguts per condition, pooled from 4 independent experiments, unpaired t-test with Welch’s correction). (G) Quantification of sporozoites isolated from salivary glands nineteen days post ookinete injections. Each data point is an average of five infected mosquitoes (representative data from n = 2 independent experiments, unpaired t-test). In all relevant panels, statistical significance is indicated as: ns = not significant, ** = p < 0.01, *** = p < 0.001 and **** = p < 0.0001.

Article Snippet: The slides were mounted using 30μl of Fluoromount-G with DAPI (Fisher Scientific) and a 40mm coverslip and left overnight to set.

Techniques: Infection, Expressing, Control, Western Blot, Immunofluorescence, Staining, Clinical Proteomics, Membrane, Marker, Isolation