<p> Supplementary Material</p><p>Journal of Nanoparticle Research</p><p>Cholesterol-conjugate supramolecular assemblies of low generations polyamidoamine dendrimers for enhanced EGFP plasmid DNA transfection</p><p>:Authors</p><p>*Nasim Golkara , Soliman Mohammadi Samania,b, Ali Mohammad Tamaddona,b</p><p>:Affiliations</p><p> a Department of Pharmaceutics, School of Pharmacy, Shiraz University of Medical Sciences, Shiraz, Iran, </p><p>71345</p><p> b Center for Nanotechnology in Drug Delivery, Shiraz University of Medical Sciences, Shiraz, Iran, 71345</p><p>*To whom correspondence should be addressed:</p><p>Dr. Ali Mohammad Tamaddon, Ph.D.</p><p>Associate Professor,</p><p>Shiraz University of Medical Sciences,</p><p>Shiraz, Iran, 71345</p><p>Telefax: (0098) 71 32424126</p><p>Email: [email protected]</p><p>1 a B</p><p>C D</p><p>E F</p><p>Fig S1 FTIR spectra of a: G2 PAMAM dendrimers; b: cholesteryl chloroformate and 40%</p><p> cholesterol conjugated G2 PAMAM dendrimer (G2-Chol40%); c: unmodified G1, G2 and</p><p>G3 PAMAM dendrimers; d, e and f: various cholesterol conjugated G1, G2 and G3</p><p>PAMAM dendrimers.</p><p>2 Table S1. Characteristic peaks in FTIR spectra of unmodified and cholesterol conjugated PAMAM dendrimers</p><p>Amine Carbonyl Amine stretching =C-H (alkene) bending stretching stretching G1 1550 cm-1 1670 cm-1 3100 cm-1 3270 cm-1 G2 1550 cm-1 1670 cm-1 3100 cm-1 3270 cm-1 G3 1550 cm-1 1670 cm-1 3100 cm-1 3270 cm-1 -1 -1 -1 -1 -1 G1-Chol15% 1550 cm 1670 cm 3050 cm 3270 cm 2900-3000 cm -1 -1 -1 -1 -1 G1-Chol40% 1550 cm 1670 cm 3050 cm 3270 cm 2900-3000 cm -1 -1 -1 -1 -1 G1-Chol60% 1550 cm 1670 cm 3050 cm 3270 cm 2900-3000 cm -1 -1 -1 -1 -1 G2-Chol15% 1550 cm 1670 cm 3050 cm 3270 cm 2900-3000 cm -1 -1 -1 -1 -1 G2-Chol40% 1550 cm 1670 cm 3050 cm 3270 cm 2900-3000 cm -1 -1 -1 -1 -1 G2-Chol60% 1550 cm 1670 cm 3050 cm 3270 cm 2900-3000 cm -1 -1 -1 -1 -1 G3-Chol6% 1550 cm 1670 cm 3050 cm 3270 cm 2900-3000 cm -1 -1 -1 -1 -1 G3-Chol15% 1550 cm 1670 cm 3050 cm 3270 cm 2900-3000 cm -1 -1 -1 -1 -1 G3-Chol40% 1550 cm 1670 cm 3050 cm 3270 cm 2900-3000 cm -1 -1 -1 -1 -1 G3-Chol60% 1550 cm 1670 cm 3050 cm 3270 cm 2900-3000 cm</p><p>3 a 0 0 0 0 1 0 0 1 2 2 0 ...... 3 3 5 8 0 9 7 8 0 5 8 9 0 0 3 4 9 6 0 4 7 9 8 1</p><p>8.0 7.0 6.0 5.0 4.0 3.0 2.0 1.0 0.0 ppm (t1) b 1 1 0 0 1 . . . . . 0 0 4 8 4 7 7 9 2 0</p><p>7.0 6.0 5.0 4.0 3.0 2.0 1.0 0.0 ppm (t1)</p><p>4 c</p><p>4 3 2</p><p>1 1 2 0 1 . . . . 3 1 4 0 2 2 0 1</p><p>7.0 6.0 5.0 4.0 3.0 2.0 1.0 0.0 ppm (t1) d 1 1 0 1 . . . . 3 5 5 0 2 5 3 0</p><p>7.0 6.0 5.0 4.0 3.0 2.0 1.0 0.0 ppm (t1)</p><p>5 e 0 0 2 1 2 1 2 5 3 5 1 ...... 7 8 6 9 6 5 9 9 5 9 0 6 4 6 0 3 1 8 9 7 3 0</p><p>7.0 6.0 5.0 4.0 3.0 2.0 1.0 0.0 ppm (t1) f 0 0 1 3 1 1 5 3 1 1 4 1 ...... 6 5 4 0 8 1 7 6 9 8 9 0 2 6 6 1 0 1 3 0 8.0 7.0 6.0 5.0 4 4.0 4 2 3.0 3 2.0 1.0 0.0</p><p>6 g 0 1 4 3 5 3 3 2 6 1 ...... 8 0 8 4 7 1 1 0 2 1 4 0 0 0 9 8 7 2 3 0</p><p>7.0 6.0 5.0 4.0 3.0 2.0 1.0 0.0 ppm (t1) h 0 0 0 2 1 1 1 2 3 1 2 1 ...... 8 2 0 4 4 1 7 8 1 6 7 3 2 4 5 6 6 7 0 4 7 9 8 0</p><p>7.0 6.0 5.0 4.0 3.0 2.0 1.0 0.0 ppm (t1)</p><p>1 Fig S2 H-NMR spectrum of a: cholesteryl chloroformate in CDCl3; b, c and d: unmodified G1,</p><p>G2 and G3 PAMAM dendrimer in D2O; e and f: 40% and 60% cholesterol conjugated G2</p><p>PAMAM dendrimer (G2-Chol40% and G2-Chol60%) in CDCl3; g and h: 40% and 60%</p><p> cholesterol conjugated G3 PAMAM dendrimer (G3-Chol40% and G3-Chol60%) in CDCl3,</p><p> respectively.</p><p>7 Table S2. Peak characteristics for 1H-NMR spectra of unmodified and cholesterol conjugated PAMAM dendrimers</p><p>Aliphatic hydrogens NH-CO- CO-NH-CH2- -CH2-NH- 14-H -C=C- (cholesterol) CH2-CH2- CH2-NH2- CO- (cholesterol) (cholesterol) G1 2.30 ppm 2.70 ppm 3.20 ppm G2 2.30 ppm 2.70 ppm 3.20 ppm G3 2.30 ppm 2.70 ppm 3.20 ppm</p><p>G1-Chol15% 0.70-2.40 ppm 2.40 ppm 2.70 ppm 3.29 ppm 4.70 ppm 5.40 ppm</p><p>G1-Chol40% 0.70-2.40 ppm 2.40 ppm 2.70 ppm 3.29 ppm 4.70 ppm 5.40 ppm</p><p>G1-Chol60% 0.70-2.40 ppm 2.40 ppm 2.70 ppm 3.29 ppm 4.70 ppm 5.40 ppm</p><p>G2-Chol15% 0.70-2.40 ppm 2.40 ppm 2.70 ppm 3.29 ppm 4.70 ppm 5.40 ppm</p><p>G2-Chol40% 0.70-2.40 ppm 2.40 ppm 2.70 ppm 3.29 ppm 4.70 ppm 5.40 ppm</p><p>G2-Chol60% 0.70-2.40 ppm 2.40 ppm 2.70 ppm 3.29 ppm 4.70 ppm 5.40 ppm</p><p>G3-Chol6% 0.70-2.40 ppm 2.40 ppm 2.70 ppm 3.29 ppm 4.70 ppm 5.40 ppm</p><p>G3-Chol15% 0.70-2.40 ppm 2.40 ppm 2.70 ppm 3.29 ppm 4.70 ppm 5.40 ppm</p><p>G3-Chol40% 0.70-2.40 ppm 2.40 ppm 2.70 ppm 3.29 ppm 4.70 ppm 5.40 ppm</p><p>G3-Chol60% 0.70-2.40 ppm 2.40 ppm 2.70 ppm 3.29 ppm 4.70 ppm 5.40 ppm</p><p>8 Fig S3 Fluorescence emission spectra of pyrene excited at 340 nm in the presence of G2-Chol40% </p><p> dispersions at various polymer concentrations (15.62 to 1000 µg/ml) in deionized water.</p><p>9</p>