Ordinary microscope

The species and genders of most mosquitoes employed in this study have been identified in our previous study [16 (link)]. The remaining samples were identified with standard morphological criteria (https://www.wrbu.si.edu/) with an ordinary microscope (Olympus, Tokyo, Japan). Each mosquito was removed from DNA/RNA stabilization reagent and placed on to a glass slide for morphological identification. In addition, the mosquitoes of each species from each province were subjected to further identification using a genomic approach targeting mitochondrial cytochrome c oxidase subunit I (COI) gene [18 (link)] (Table 1). After species and gender identification, the mosquitoes were stored at -80°C until genomic DNA extraction.

Human colon cancer cell line: SW620, CW-2, LoVo and HCT116 purchased from ATCC (90% DMEM + 10% FBS) 0.1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethyleneglycol)-2000(ammonium salt) (DSPE-PEG2000-NH2) were purchased from Ananti Polar Lipids, Inc. (Beijing, China). Cetuximab, a monoclonal antibody anti-EGFR, was purchased from Shanghai TheraMabsBiotech (product number TM-Beva-00002). DMSO (D8371, Solarbio), dialysis bag (3.5KD, F132590, BBI), DMEM medium (Cat. No. 11965-126), fetal bovine serum (FBS) (product number 16000-044), trypsin and double antibodies were purchased from Gibco company. MTT powder was purchased from Azerbaijan Latin (Cat. No. T100896-1g), irinotecan (Irinotecan, CPT-11), purchased from Solarbio (SI8480-20 mg), EDC purchased from Sigma (D8740-500 mg), NHS purchased from Sigma (D8740-500 mg) Balb/c nude mice were purchased from Weitong Lihua.
Main instruments: ordinary microscope (OLYMPUS company in Japan), small desktop refrigerated centrifuge (Technology University Zhongjia), cell incubator (American Thermoelectric), heating magnetic stirrer (C-MAG HS 10, IKA) CM-H₂DCFDA were obtained from ThermoFisher.

For wound healing assays, MCF-7 cells (2.8 × 10⁵ cells per well) were seeded into 6-well plates and treated with cRGD-CL/pshOC-2 lipoplexes (pshOC-2 = 1.5 μg/well, N/P ratios = 2, 3 and 4), bare liposomes cRGD-CL and CL, respectively, in Opti-MEM for 24 h. Then cells were wounded using pipette tips and cell migration was imaged at 0 h and 24 h using an inverted microscope (Carl Zeiss, Thuringia, Germany).
For Transwell migration assay, MCF-7 cells were treated with cRGD-CL/pshOC-2 lipoplexes (pshOC-2 = 1.5 μg/well, N/P = 4), cRGD-CL and CL for 24 h, respectively, and then seeded into the Transwell chamber (8 μm) at 8 × 10⁴ cells per chamber and incubated for another 24 h. Cells were fixed with 4% paraformaldehyde, stained with crystal violet, and imaged with an ordinary microscope (Olympus).