MOESM1 of Infectious salmon anaemia virus (ISAV) mucosal infection in Atlantic salmon

Additional file 1. Statistical modelling of 4 immune marker expression profiles. Statistical modelling of 4 immune marker expression profiles, type I IFN (A), Mx (B), type II IFN (C) and ÎłIP (D) in hind gut of fish infected with high virulent (dashed line and triangles) or low virulent (solid line and circles) strains measured by real-time RT-qPCR. Dotted lines indicate 95% confidence intervals

RT-qPCR of gills and gill swabs, and immunostaining on frozen gills from farm IV.

<p>RT-qPCR of gills and gill swabs, and immunostaining on frozen gills from farm IV.</p

Immunostaining of tissue sections.

<p>Immunostaining of formalin-fixed paraffin-embedded (A-C), and frozen (D-I) gill sections with massive and sparse infection ISAV HPR0 infection, demonstrating epithelial infection. Primary antibodies used were a rabbit antibody to ISAV NP (A-D, G), and a monoclonal antibody to ISAV HE (E-F, H-I). For detection of bound antibodies, alkaline phosphatase-conjugated antibody to rabbit Ig with Fast Red substrate (A-D; red colour), horseradish peroxidase-conjugated antibody to mouse Ig with DAB substrate (E-F; brown colour), and Alexa fluor 488-conjugated antibody to rabbit Ig (G) or mouse Ig (H-I), were used.</p

Summary of RT-qPCR on gills.

<p>Summary of RT-qPCR on gills.</p

Mx and γIP response to HPR0 infection at farm III.

<p>Across the three months, little variation in Mx expression (A) was seen. However, γIP was over 8-fold lower in the fish sampled in August compared to those sampled in September and October (B), suggesting the presence of a γIFN-induced response. In August 2009, all fish (n = 41) tested negative (neg.) for HPR0. In September 2009, 72% (58 out of 80) fish tested positive (pos.) (mean Ct 31.5, range 20.8–36.7). In October 2009, 5% (4 out of 79) fish tested positive (mean Ct 35.6, range 34.2–36.5). Ratios standardised relative to elongation factor 1α (ELF).</p

RT-qPCR on gills and fin swabs.

<p>RT-qPCR on gills and fin swabs.</p

Immune marker RT-qPCR.

<p>Immune marker RT-qPCR.</p

Virus susceptibility of five cell lines expressed as the calculated log₁₀ TCID₅₀ titer per 50 μl of a stock culture of six salmonid viruses.

<p>Virus susceptibility of five cell lines expressed as the calculated log₁₀ TCID₅₀ titer per 50 μl of a stock culture of six salmonid viruses.</p

Measurement of scratch width.

<p>Relative measurement of the width of the scratch in cm to assess repair abilities of the two cell lines. The distance of the scratch was measured in cm on enhanced snapshots. The plot displays the width of the scratch over time.</p

Cell culture migratory and proliferative properties.

<p>Pictures were taken daily during a 7 days scratch closure assay. Representative pictures of ASG-10 (a-d) and ASG-13 (e-h) at day 0 (a and e), day 3 (b and f), day 6 (c and g) and day 7 (d and h) are shown.</p