Fig 2: Effect of DON treated with SMBS or encapsulated SMBS (ES) on the gene and protein expression of tight junctions in IPEC-J2 cells. Note: IPEC-J2 cells were cultured in 12-well plates (for gene expression) or 6-well plates (for protein expression) until 90% confluent and then incubated with DON (1 µg/mL) or the same concentration of DON treated with SMBS (0.5%) or ES (containing 0.5% SMBS) (2 h) as described in the Materials and Methods section for 4 h (for gene expression) or 24 h (for protein expression). Total RNA and protein were extracted. Gene expression of zonula occludens-1 (ZO-1) (A) and occludin (OCLN) (B) was measured by RT-PCR, and data were expressed as fold change relative to control (CON) and presented as mean ± SD, n = 4. Different letters indicate significant difference (P < 0.05). Protein expression of ZO-1 (D) and OCLN (E) was detected by Western blot analysis, and representative images (C) were presented. CON: control; D + S: DON treated with SMBS; D + ES: DON treated with encapsulated SMBS.

Fig 3: Neural stem cells in the axolotl spinal cord reactivate molecular features associated with embryonic neuroepithelial cells upon injury.(A) Schematic of the distinct regions microdissected for NanoString expression profiling. 48–50 pieces of spinal cord were pooled together for each of the 3 biological replicates per group. (B) Changes in gene expression (log2 fold change) in cells of the source zone at day 1 (left) and regenerating spinal cord at day 6 (right) versus cells in the uninjured spinal cord (day 0). Y-axis shows the statistical significance of these changes (-log10 p value). Red dots, upregulated genes; blue dots, downregulated genes; grey dots, genes that are not significantly regulated in regenerating versus uninjured samples. N=3 samples per group. P values indicate the results of negative binomial tests. p<0.05 (FDR < 0.07) was considered significant. Dot area is proportional to the number of transcripts counted for that gene. Names are the top five upregulated (red) and top five downregulated (blue) genes in the dataset. (C) Heat map of fold change in gene expression values in regenerating spinal cord versus uninjured spinal cord, side by side with the reported fold change values in the stem zone and pre-neural tube (SZ+PNT) compared to more advanced rostral and caudal neural tube (CNT+RNT) of the embryonic chick spinal cord, as described in (Olivera-Martinez et al., 2014). Dataset used to generate the volcano plots and heat maps can be found in Figure 1—source data 1. (D) In situ hybridization on spinal cord cross-sections at day 0 (top) and of the regeneration source zone at day 1 (bottom). (E) In situ hybridization on spinal cord cross-sections at day 0 (top) and regenerating spinal cord at day 6 (bottom). (F) Immunofluorescence on cross-sections of uninjured (top) and regenerating spinal cord at day 6 (bottom) for the apical junctional proteins ZO-1, OCLN, and CLDN5. DNA is labeled with Hoechst. Insets show the apical surface of the spinal cord at higher magnification. Images are single optical sections using the same acquisition parameters on sections from the same slide. Observations were consistent in more than 5 axolotls. Scale bars, 20 µm.DOI: http://dx.doi.org/10.7554/eLife.10230.00310.7554/eLife.10230.004Figure 1—source data 1Gene expression changes in the regenerating axolotl spinal cord compared to the uninjured axolotl spinal cord.DOI: http://dx.doi.org/10.7554/eLife.10230.004

Fig 4: HIV-1 infection and/or modifications of the cav-1/ocln/Alix complex affect anti- and pro-inflammatory cytokine profile in pericytes. Pericytes were transfected with cav-1 siRNA (Cav-1-), ocln expression vector (Ocln+), or Alix siRNA (Alix-) as in Figure 5 and either mock-infected or HIV-1-infected for 48 or 72 h. Cytokine levels were analyzed in cell culture media by Bio-Plex Pro Human Cytokine assay kit. INF-? levels decreased to non-detectable (#) concentrations after HIV-1 infection in control, cav-1-, and Alix- pericyte cultures. The data are mean ± SEM from two independent experiments. ****P < .0001, ***P = .0002, **P = .003, *P < .0449, n = 4 per group

Fig 5: Organ-specific localization of OCLN, JAM-A and ZO-1 in canine duodenum, lung, liver, and kidney, as detected by immunohistochemistry. (A) OCLN, (B) JAM-A and (C) ZO-1 immunostaining was detected in all organs examined. Boxes in the upper panels (magnification, ×200) are areas magnified in the lower panels (magnification, ×400). Arrows indicate positive staining. OCLN, occludin; JAM-A, junction adhesion molecule A; ZO-1, zona occludens 1.