Human tubulin beta 2A class IIa, Tubb2a For: gagggcgaggacgaggctta 122 (NM_001069.2) Rev: tctaacagaggcaaaactgagcacc Supplementary Figures Fig. S1. Integrity and size of the chemically modified mRNAs as determined by native agarose gel electrophoresis. cmRNAs and RiboRuler RNA ladder high range were mixed with RiboRuler formamide containing loading dye and incubated for 10 minutes at 70 °C. Subsequently samples were chilled on ice and applied to the agarose gel. Detection was done by ethidiumbromide staining and visualization on Intas Gel Imaging System. Fig. S2. luciferase expression of transfected and cytotoxicity associated to Metridia (A) Kinetics of the luciferase expression in the used MetLuc cmRNA complexes. Metridia cells transfected with complexes formed using Lipofectamine2000 (LF), Dogtor, DF-Gold or bPEI as enhancers at a cmRNA dose of 20 pg/cell. n=3, One-way ANOVA. Tukey’s multiple comparison test. (B) Viability of the cells 5 and 24 hours after transfection. Significant difference between untransfected control (100% cell viability) and transfected cells are indicated by (*). n=3, Student’s t-Test, 2 tailed. Range of Fig. S3. Titration of DF-Gold and MetLuc cmRNA amounts in AMSCs. tested was from 0.5 to 5 per nucleic acid. The amount of MetLuc cmRNA used μl μl μg for the titration was 2.5, 5, 10 and 20 pg/cell. (A-F) 5, 24, 48, 72, 168 and 240 hours
Human tubulin beta 2A class IIa, Tubb2a (NM_001069.2) For: gagggcgaggacgaggctta Rev: tctaacagaggcaaaactgagcacc 122 Supplementary Figures Fig. S1. Integrity and size of the chemically modified mRNAs as determined by native agarose gel electrophoresis. cmRNAs and RiboRuler RNA ladder high range were mixed with RiboRuler formamide containing loading dye and incubated for 10 minutes at 70 °C. Subsequently samples were chilled on ice and applied to the agarose gel. Detection was done by ethidiumbromide staining and visualization on Intas Gel Imaging System. Fig. S2. Metridia luciferase expression of transfected and cytotoxicity associated to the used MetLuc cmRNA complexes. (A) Kinetics of the Metridia luciferase expression in cells transfected with complexes formed using Lipofectamine2000 (LF), Dogtor, DF-Gold or bPEI as enhancers at a cmRNA dose of 20 pg/cell. n=3, One-way ANOVA. Tukey’s multiple comparison test. (B) Viability of the cells 5 and 24 hours after transfection. Significant difference between untransfected control (100% cell viability) and transfected cells are indicated by (*). n=3, Student’s t-Test, 2 tailed. Fig. S3. Titration of DF-Gold and MetLuc cmRNA amounts in AMSCs. Range of tested was from 0.5 μl to 5 μl per μg nucleic acid. The amount of MetLuc cmRNA used for the titration was 2.5, 5, 10 and 20 pg/cell. (A-F) 5, 24, 48, 72, 168 and 240 hours