Fig. 7

Expression of COL-99::EGFP::FLAG in C. elegans. a Verification of the transgenic worm line col-99::egfp::flag by RT-PCR. Primers are specific for cDNA of EGFP, col-99 and the reference gene tba-1 respectively. b Western blot analysis of the EGFP- and FLAG-tagged COL-99 protein in C. elegans lysates resolved under reducing conditions. Lanes from left to right are lysates from worm lines unc-119, col-99::egfp::flag, and from CHO cell expressing human collagen XIII with an EGFP-tag. β-tubulin was used as a loading control. c Western blot of col-99::egfp::flag worm lysate with or without DTT reducing agent treatment. Arrows indicate the COL-99::EGFP::FLAG protein in monomeric (~120 kDa) or trimeric (>300 kDa) forms. The detection antibody was anti-FLAG. d Localization of COL-99::EGFP::FLAG at the worm body wall (BW) imaged by confocal microscopy. e Green fluorescence imaging of the control line unc-119. The asterisks in (d) and (e) indicate autofluorescence produced by the bacteria in the C. elegans intestine. f In vivo imaging of an adult worm mouth (MO) part. The 2D image was converted from a 3D confocal scanning. g Image of the same region in a control unc-119 worm. h Whole worm imaging of a L1 larva (HE = head, TA = tail). i Control image of an unc-119 L1 larva. Bars in (d)-(i), 10 μm. About ten worms of each type were imaged in total