Journal: PLOS Neglected Tropical Diseases

Article Title: Giardia duodenalis MIF induces host intestinal damage via CD74 receptor mediated NLRP3 inflammasome activation

doi: 10.1371/journal.pntd.0013968

Figure Lengend Snippet: 1 × 10 7 trophozoites were incubated with PBS, NC antibody or GdMIF antibody for 1 h, respectively, then trophozoites were used to infect gerbils, all gerbils were euthanized and collected duodenum at 10 dpi (n = 5 per group). (A) The number of trophozoites in the duodenal lavage fluid were counted. (B) The mRNA expression levels of cytokines in the duodenum were detected by qPCR. (C) Representative images of H&E staining in the duodenum. The blue arrow points to G. duodenalis , scale bar: 50 μm and 200 μm. Four images were captured in total per sample (n = 5), and the villus length and crypt depth were measured as indicated in the image. (D) The statistical analysis of villus length, crypt depth, and the ratio of the villus to crypt of the duodenum. (E) Histopathological score of duodenum. (F) AB-PAS staining of goblet cells in duodenum tissue, MUC2 and ZO-1 expression in duodenum tissue was observed via IHC, scale bar: 100 μm. Four images were captured in total per sample (n = 5). (G) The number of goblet cells in the image was analysed using Imag J (n = 5). (H) The level of MUC2 in the image was quantified using Imag J. (I) The level of ZO-1 in the image was quantified using Imag J. (J-M) The expression levels of ZO-1, Occludin, and Claudin3 proteins in duodenum were detected by western blot and densitometric analysis (n = 3). All results were representative of three number of biologically independent experiments. Data was shown as the mean ± SD, statistical significance was assessed using unpaired Student’s t-test. n.s. p > 0.05 indicates not significant, * p < 0.05, ** p < 0.01, *** p < 0.001 and **** p < 0.0001.

Article Snippet: Tissue sections were deparaffinized in xylene, rehydrated through a graded ethanol series, and stained using the AB-PAS stain kit (Cat. No. G1049, Servicebio) as per the manufacturer’s instructions.

Techniques: Incubation, Expressing, Staining, Western Blot

Journal: PLOS Neglected Tropical Diseases

Article Title: Giardia duodenalis MIF induces host intestinal damage via CD74 receptor mediated NLRP3 inflammasome activation

doi: 10.1371/journal.pntd.0013968

Figure Lengend Snippet: (A) Gerbils were administrated 1 × 10 7 trophozoites or PBS for 3h, then gerbils were received MCC950 (10 mg/kg) i.p. daily for 10 days in the MCC950 inhibitor group. (B) The body weights of gerbils were monitored for 10 dpi (n = 5 per group). (C) F4/80 and NLRP3 expression in duodenum were observed by IFA. The nuclei were stained with hoechst (blue), F4/80 (green) and NLRP3 (red). Scale bar: 100 μm. Four images were analyzed in total per sample (n = 5). The positive area of F4/80 and NLRP3 in duodenum image of per sample (n = 5) was quantified using Imag J. (D) Representative images of H&E staining in the duodenum. The blue arrow points to G. duodenalis , scale bar: 50 μm and 200 μm. Four images were captured in total per sample (n = 5), and the villus length and crypt depth were measured as indicated in the image. (E) The statistical analysis of villus length, crypt depth, and the ratio of the villus to crypt of the duodenum. (F) Histopathological score of duodenum. (G) AB-PAS staining of goblet cells in duodenum tissue, MUC2 and ZO-1 expression in duodenum tissue was observed via IHC, scale bar: 100 μm. Four images were captured in total per sample (n = 5). (H) The number of goblet cells in the image was analysed using Imag J (n = 5). (I) The level of MUC2 in the image was quantified using Imag J. (J) The level of ZO-1 in the image was quantified using Imag J. (K-N) The expression levels of ZO-1, Occludin, and Claudin3 proteins in duodenum were detected by western blot and densitometric analysis (n = 3). All results were representative of three number of biologically independent experiments. Data was shown as the mean ± SD, statistical significance was assessed using using one-way ANOVA and unpaired Student’s t-test. n.s. p > 0.05 indicates not significant, * p < 0.05, ** p < 0.01, *** p < 0.001 and **** p < 0.0001.

Article Snippet: Tissue sections were deparaffinized in xylene, rehydrated through a graded ethanol series, and stained using the AB-PAS stain kit (Cat. No. G1049, Servicebio) as per the manufacturer’s instructions.

Techniques: Expressing, Staining, Western Blot