Additional file 1: of Efficient and tumor-specific knockdown of MTDH gene attenuates paclitaxel resistance of breast cancer cells both in vivo and in vitro

Table S1. Characteristics of 44 breast cancer patients with neoadjuvant chemotherapy. Figure S1. The MTDH mRNA and protein expressions level in different groups after transfecting. We constructed four groups of shRNA and one control shRNA. Then we selected the optimal silent shRNA via real-time PCR and western blot. Figure S2. Annexin V-PE/7-AAD assay for determination of apoptosis of cells overexpressing or knocking down metadherin (MTDH) with a flow cytometer. Annexin-positive cells are presented in gate 4. Figure S3. Annexin V-PE/7-AAD assay for the determination of apoptosis of different cells after paclitaxel (TAX) treatment. The apoptosis rate of MCF-7–metadherin–short hairpin RNA (MCF-7–MTDH–shRNA) cells was significantly enhanced. Figure S4. Flow cytometry was adopted to analyze cell cycle after cells overexpressing or knocking-downing MTDH. Compared to MCF-7 cell, the MCF-7-MTDH cell had more S phase and less G0/G1 and G2/M phases, while knockdown of MTDH did the opposite. Figure S5. Cell cycle assay for different cells after paclitaxel (TAX) treatment. Compared with MCF-7 and MCF-7-vector, the G2/M phase rate of MCF-7–metadherin–short hairpin RNA (MCF-7–MTDH–shRNA) cells was significantly enhanced. While overexpression of MTDH did the opposite. Figure S6. Paclitaxel (TAX) release from the polymer nanoparticles (NPs). The NPs showed a faster release rate for TAX over time in PBS at pH 4.4 than at pH 7.4. Each bar represents the mean ± standard deviation of three replicates. Figure S7. In vivo tumor targeting of nanoparticles (NPs). Nude mice bearing MCF-7 tumors (~100 mm3) were given a single intravenous injection of Cy5.5-labeled free small interfering RNA (siRNA) or NP-TAX–siRNA by the tail vein. In vivo fluorescence signals were recorded by using a Maestro2.10.0 imaging system for up to 24 h post-injection. Abbreviation: TAX paclitaxel. (DOC 2597 kb