Supporting Table. the Half Maximal Inhibitory Concentration (IC50) of PTX-Loaded Nps
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Supporting information – Gullotti et al.
Supporting Table. The half maximal inhibitory concentration (IC50) of PTX-loaded NPs.
PTX/PLGA-pDA-TAT NPs PTX/PLGA NPs log(inhibitor) vs. response (three parameters) Best-fit values Bottom 29.18 15.37 Top 92.92 101.6 LogIC50 1.861 2.413 IC50 72.60 258.7 Span 63.75 86.28 PLGA NPs NP Nu
Tm Overlay
PLGA-pDA NPs
PLGA-pDA-TAT NPs
Supporting Fig. 1. Cellular uptake of *PLGA, *PLGA-pDA, and *PLGA-pDA-TAT NPs. NPs were added to SKOV-3 cells in serum-free medium. NP: green fluorescence signal from NPs; Nu: nuclei stained with Draq5; Tm: transmission image; Overlay: overlay of NP, Nu and Tm. Magnification: 600×. Scale bar = 40 μm. Supporting information – Gullotti et al.
Primers MMP-2 MMP-9 GAPDH cDNA OV SK OV SK OV SK
Supporting Fig. 2. Semi-quantitative RT-PCR. A low level of MMP-9 mRNA production was observed in both SKOV-3 (SK) and OVCAR-3 (OV) cell lines. MMP-2 mRNA levels in SKOV- 3 cells were higher than in OVCAR-3 cells. GAPDH was used as a house-keeping gene. Experimental: mRNA was extracted from OVCAR-3 and SKOV-3 cells using an RNeasy extraction kit (Qiagen). cDNA synthesis from mRNA was done by SuperScriptTM (Invitrogen) according to the manufacturer’s protocol. Semi-quantitative PCR was performed using an Eppendorf thermal cycler. The following primers were used in a standard PCR reaction: MMP2 (F)- 5’ CGG CCG CAG TGA CGG AAA 3’; MMP2 (R)- 5’ CAT CCT GGG ACA GAC GGA AG 3’; MMP9 (F)- 5’ GGA AGA TGC TGC TGT TCA GC 3’; MMP9 (R)- 5’ ACC TGG TTC AAC TCA CTC CG 3’; GAPDH (F)- 5’ AGA AGG CTG GGG CTC ATT TG 3’; GAPDH (R)- 5’ AGG GGC CAT CCA CAG TCT TC 3’. Reaction conditions: 94°C for 2 min followed by 25 cycles (94°C for 30 sec, 55°C for 30 sec, 72°C for 1 min) and final extension at 72°C for 1 min. PCR products were then resolved on a 1.25% agarose gel containing ethidium bromide and visualized with UV light. The relative intensities of the bands were analyzed qualitatively.