Mircury lna target site blocker

Following 4 weeks of HFD feeding, Apoe^−/− mice were randomized to the different experimental groups and injected once weekly via the tail vein with KLF4 LNA-target site blocker oligonucleotides (5′- GTATGCAGCAGTTGG -3′) or control LNA-target site blocker oligonucleotides (5′- GCTCCCTTCAATCCAA -3′; 0.4 mg per 20 kg; miRCURY LNA Target Site Blocker, in vivo use; Exiqon). During the injection period, mice were fed with a HFD. The tissues were harvested 1 week after the last injection.

Following 8 weeks of HFD feeding, Apoe^−/− mice (The Jackson Laboratory, Bar Harbor, ME, USA) were randomized to the different experimental groups and tail vein injected once weekly for four consecutive weeks with lncWDR59 LNA-TSB or control LNA oligonucleotides (0.5 mg per 20 g; miRCURY LNA Target Site Blocker, in vivo use; Exiqon). During the injection period, mice were fed with a HFD comprising 21% crude fat, 0.15% cholesterol, and 19.5% protein. Tissues were harvested 1 week after the last injection. Paxgene-fixed/paraffin-embedded aortic roots were stained with Elastic van Gieson stain. A bright-field microscope (Leica DM6000B; Leica Microsystems) connected to a CCD camera (Leica DFC365) was used to obtain the images. The lesion area was quantified with ImageJ. Immunostaining of Ki67, γH2AX, C-X-C motif chemokine ligand 1 (Cxcl1), and von Willebrand factor (vWF) was also performed. The positive area or the number of positive cells was normalized to the lesion area or to the total number of vWF⁺ cells, respectively, using ImageJ. The analysis of the staining was performed in a blinded manner.
All animal experiments were reviewed and approved by the local authorities (State Agency for Nature, Environment and Consumer Protection of North Rhine-Westphalia and District Government of Upper Bavaria) in accordance with the German animal protection laws.