In vivo jet pei delivery reagent

The mouse EM model was established by endometrial auto-transplantation according to previous description [21 ]. Briefly, all mice were anesthetized by intraperitoneal injection of 1% pentobarbital sodium (40 mg/kg). Then, a small midline incision was made on the abdomen, and the left uterine horn was excised and immediately placed in a physiological saline solution. The endometrium was carefully separated from muscles and cut into two segments (5 mm × 5 mm). A subcutaneous pocket was created on each side of the abdominal wall and the uterine segments were placed in the spaces with the endometrium facing the abdominal muscle. Then, the abdominal incision was sutured. One day after the surgery, the mice were randomly divided into two groups (n = 6/group). The mice were injected intraperitoneally with 90 μg of NC inhibitor or miR-429 inhibitor and 20 μL of in vivo‐jet PEI delivery reagent (Polyplus Transfection, Illkirch, France) in 5% glucose every 2 days, respectively. After 2 weeks, the mice were euthanized under anesthesia, and endometriotic lesions were collected for subsequent experiments. All results were assessed by two investigators blinded to the group treatments.

BALB/c or C57BL/6 mice were IV tail-injected with 250 μl of the in vivo-jet PEI delivery reagent (Polyplus transfection, France) complexed with an appropriate luc reporter plasmid. 24 hours later the mice were injected with 250 μl of 3 mg/ml luciferin solution (Gold Biotechnology, USA) and underwent bioluminescence imaging by Biospace Photon Imager (Biospace Lab, France).

Specific-pathogen-free, six-week-old golden Syrian hamsters were purchased form Janvier Labs, and they were kept in individually ventilated cages. Hamsters were inoculated intranasally with 4×10⁵ PFU of SARS-CoV-2 virus. In vivo transfection was performed as previously described (26). Briefly, plasmid or siRNA was combined with in vivo-jetPEI delivery reagent (Polyplus-transfection, NY, USA) in a 10% glucose solution according to the manufacturer’s instruction. The solution was mixed and incubated for 30 min at room temperature, and then were intravenous injected into hamsters 12 hours before SARS-CoV-2 infection. Strengthen injection was performed at 3 dpi. Knockdown or overexpression efficiency was detected by western blotting at 2, 4 and 7 dpi.