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Figure 1

From: C. elegans expressing D76N β2-microglobulin: a model for in vivo screening of drug candidates targeting amyloidosis

Figure 1

D76N β2-m C. elegans strain genotype characterization and β2-microglobulin expression. (a) Scheme of C. elegans manipulation highlighting the stage in which the β2-m expression is activated by temperature switch. Eggs were maintained at 16 °C and L1 larvae were up-shifted to 23–25 °C in order to induce the expression of the protein. (b) PCR genotyping of adult transgenic nematodes. The expected size of PCR products (about 800 bp) was observed after DNA electrophoresis on 1.5% agarose gel (lane 1: EZ Load Precision, BioRad, lane 2,4: pcr products of DNA extracted from smg-1 (cc546) ancestral strain, lane 3,5: pcr products of DNA extracted from D76N β2-m worms. (c) Representative western blot of β2-m expression. Equal amounts of protein (10 μg) were loaded for each sample from smg-1 (-ctrl) or D76N β2-m expressing nematodes and immunoblotted with polyclonal anti-human β2-m antibody (Dako) and anti-GAPDH antibody used as loading control (M = Molecular weight standard: Precision Plus Western C, BioRad). Uncropped scans of immunoblots are shown in Supplementary Fig. S1.

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