Hoechst 33258

Polyamidoamine dendrimer (G4-PAMAM, MW = 14,214 Da) was purchased from Sigma-Aldrich (St. Louis, MO, USA). Hyaluronic acid was purchased from Shandong Freda Biotech Co., Ltd. (Shandong, China). TRNzol A+ reagent was purchased from Tiangen (Beijing, China). Lipofectamine 2000 was purchased from Invitrogen (Carlsbad, CA, USA) The scrambled siRNAs (siN.C.), the FAM-labeled negative siRNA (FAM-siRNA) (antisense strand, 5′-ACGUGACACGUUCGGAGAATT-3′), and siRNA anti-GFP (siGFP, antisense strand, 5′-GAUCUCAUCAGGGUACUCCdTdT-3′) were purchased from Genepharma (Shanghai, China). The siRNAs were double-stranded RNA oligomer containing 21 nt. All primers were synthesized by AuGCT Biotechnology (Beijing, China). RPMI-1640 medium, modified eagle medium (MEM), Dulbecco’s MEM (DMEM), penicillin-streptomycin, trypsin, and Hoechst 33258 were purchased from Macgene Technology (Beijing, China). LysoTracker Red and Green was purchased from Invitrogen (Carlsbad, CA, USA). The reverse transcription system was purchased from Promega (Madison, WI, USA). Other reagents were commercially available and used without further purification.

Distearoyl-glycerophosphoethanolamine-polyethyleneglycol₂₀₀₀-maleimide (DSPE-PEG₂₀₀₀-Mal) was purchased from NOF Corporation (Tokyo, Japan). Peptide (CYKLEGTTRLTRKRGLKLA) was custom synthesized by GL Biochem (Shanghai, China). Paclitaxel (PTX) was purchased from Norzer Pharmaceutical Co., Ltd. Alpha linolenic acid (ALA) was purchased from ANPEL Laboratory Technologies (Shanghai, China). Soybean phosphatidylcholine (SPC), unesterified cholesterol (UC), triolein (TO) and cholesteryl oleate (CO) were from Lipoid (Ludwigshafen, Germany), Wako Ltd. (Tokyo, Japan), Sinopharm Chemical Reagent Co., Ltd (Beijing, China) and InnoChem Science & Technology Co., Ltd. (Beijing, China), respectively. Solutol^® HS 15 was produced by BASF (Germany). Dicyclohexylcarbodiimide (DCC) and 4-dimethylaminopyridine (DMAP) were obtained from Sinopharm Group Co., Ltd. Cell Counting Kit-8 (CCK- 8) was obtained from SolarbioScience & Technology Co., Ltd (Beijing, China). Coumarin-6 (COU) was purchased from Sigma-Aldrich Company (USA). TUNEL Kit was obtained from KeyGEN Biotech Co., Ltd. (Nanjing, China). Enzyme-linked Immunosorbent Assay (ELISA) Kit for LDLR was obtained from USCN Life Science Inc. Modified eagle medium (MEM), DMEM, penicillin-streptomycin, trypsin and Hoechst 33258 were obtained from Macgene Technology (Beijing, China).

An intracellular replication assay was performed to determine the number of parasites per vacuole at 24 h post-invasion. HFFs growing in 12-well plates seeded on coverslips were inoculated with 1 × 10⁵ parasites. After 1 h, uninvaded parasites were removed and cultured for 24 h with IAA (500 μM) or a vehicle (Ethyl Alcohol, 1:1000). The parasites were then fixed and co-stained with mouse anti-HA (diluted to 1:500, Sigma, St. Louis, MO, USA) and rabbit anti-αGAP45 polyclonal antibodies (diluted to 1:300, a gift from Professor Qun Liu at the Chinese Agricultural University). Nuclear DNA was stained with Hoechst 33258 (diluted to 1:100, Macgene, Beijing, China). Parasites of each strain in vacuoles were quantified by counting at least 100 vacuoles using a fluorescence microscope (Zeiss, Oberkochen, Germany). Three independent experiments were performed. For the invasion assay, the percentage of invasion was calculated based on the number of vacuoles per host cell. Three independent experiments were conducted.