Alzet micro osmotic pumps model 1007d

The antisense oligonucleotide used in this study was fully described before (Michailidou et al., 2018 (link)). The sequence of the C6 antisense oligonucleotide is 5′-AACttgctgggAAT-3′ [Locked Nucleic Acid (LNA) in capital letters, DNA in lowercase letters]. This C6 LNA oligonucleotide (RGS1104) referred to as C6 antisense, targeting the mRNA of the complement component C6, was synthesized with phosphorothioate backbones and methylated DNA-C (medC) by Ribotask (Odense, Denmark), on a Mermade 12, using 2 g NittoPhase (BioAutomation). A control oligonucleotide (5′-ATCttcgcgtgaaTAA-3′) was also synthesized by Ribotask using the same method. Throughout the process, the oligonucleotide constitution was confirmed by MALDI-TOF mass spectrometry analysis on a Bruker Autoflex using 3-hydroxypicolinic acid as matrix. The antisense oligonucleotides (5 mg/kg) were dissolved in phosphate buffered saline (PBS, pH 7.4, Life technologies, Bleiswijk, The Netherlands) when administered to the mice. The antisense oligonucleotides were administered subcutaneously (s.c.) either by injection or through osmotic minipumps (Alzet micro-osmotic pumps model 1007D, Durect Corporation, Cupertino, CA, USA) as indicated for each experiment.