Fig. 11

Representative images of immunolabelling background controls. A. Salivary apparatus of a C. sonorensis orally fed on BTV-spiked blood, but BTV negative. B. Salivary apparatus of a C. sonorensis fed on mock blood and, C. Salivary apparatus of a C. sonorensis mock infected by intrathoracic inoculation. In A and B cell nuclei are shown in blue (DAPI staining), and cellular tubulin is visualised in red (labelling with mouse anti-tubulin, and anti-mouse IgG AlexaFluor™ 568). Absence of green denotes absence of BTV structural proteins (VSPs) which were labelled with in-house antibody Orab279 and anti-guinea pig IgG AlexaFluor™ 488. In panel C, cellular tubulin is shown in blue (labelling with mouse anti-tubulin, and anti-mouse IgG AlexaFluor™ 405). Absence of red or green denotes absence of BTV non-structural protein 2 (NS2) or BTV VSPs, respectively. Viral protein NS2 was labelled with in-house rabbit antibody Orab1 and anti-rabbit IgG AlexaFluor™ 568; and viral VSPs were labelled with Orab279 followed by anti-guinea pig IgG AlexaFluor™ 488 as in panels A and B. In A, one main salivary gland (SG) with its respective four accessory glands (1 to 4) and the salivary duct (arrow) can be observed. All panels are maximum projections of at least 50 stacked plane images. Microscope magnification is shown on the top right corner and the scale bar represents 20 µm