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Western-blot analysis was performed using primary antibody against soybean leghemoglobin GmLba (a generous gift from Professor Yangrong Cao of Huazhong Agricultural University, China) and HRP-conjugated rabbit anti- goat secondary antibody. As a positive control, Glutathione S-transferase (GST) tagged LjLb3 was expressed in E. coli BL21.Cells were disrupted by high pressure homogenization (D-3L ; PhD Technology International, MN, USA) and protein was puriied by GST column following recommended procedures (Sangon Biotech, China).

The luminescent Ln–ONFC ethanol suspensions were poured into the extruder (NanoAble-150, PhD Technology LLC, USA) to remove ethanol with the controlled pressure of 0.6 MPa under N2 gas, and nanopaper was formed on the lter membrane (PC, nuclepore track-etch membrane, pore size 100 nm, Whatman, UK) in less than 15 min. Then, the nanopaper was peeled o? and sandwiched between two pieces of glass and dried in ambient atmosphere.

To avoid clogging during homogenization, the suspension was first treated with ultrasonic (JY92-IIN, Scientz) for 10 min at a power of 500 W followed by high-shear mulser shearing for 1 min at 10,000 rpm (I25, IKN Co., Ltd, Shanghai, China) to obtain a homogeneous cellulose suspension. Then, the suspension was homogenized (D-3L, PhD Tech- nology, St. Paul, MN, USA) to obtain the final PCNF suspension.

These crude emulsions were homoge- nized using high-pressure homogenizer (D-3 L, PhD Technology LLC, USA) at 60 MPa for 3 cycles to generate the primary Pickering emulsion. Volumes of 45 mL, 40 mL, or 30 mL of polyphenol solutions were slowly added to achieve 5, 10, or 20 % (v/v) oil-phase ratio, respectively. The mixtures were homogenized in high-speed homogenizer (10,000 rpm, 1 min), followed by 3 additional cycles in the high-pressure homogenizer at the previously mentioned conditions.

Albumin encapsulated Ce6/ECNTs (ACEC) were prepared as follows. Brie?y, 160 mg albumin was dissolved in 10 mL ultrapure water and encapsulated into 40 mg Ce6 under high pressure ho- mogenization (PhD Technology International, MN, USA) at 4 O C. The product was puri?ed by dialysis (cutoff ? 100 kDa) against ultra- pure water at room temperature for 24h.

MnO2 dissolved in trichloromethane with IHANP in ultrapure water and homogenized (D-3L, PhD Technology LLC, MN, USA) for 15e20 min (20 000e25 000 psi). The resulting mixture was added into the ultra?ltration tube (Cut off ? 100 kDa) and was further centrifuged at 8000 rpm for 30 min to remove free MnO2, then freeze dried.

Next, 80 mg of HA-CA conjugates were dissolved in 14mL of distilled water and subjected to a high-pressure homogenizer (D-3L, PhD Technology) for 5 min to form HA nanoparticles (HANPs). Hydrophobic GKT 831 was loaded to HANP via a high-pressure homogenizer at a ratio of 10 mg GKT831/40 mg of HANP. Thirty milligrams of GKT831 (MedKoo Biosciences #522357) dissolved in 1mL of DMSO was added dropwise into the solution containing 120 mg of HANP and then circulated for an additional 5 min in the homogenizer.