D luciferin sodium salt

Male C57 mice aged 6–8 weeks were purchased from the Laboratory Animal Center of the Shandong University. The MB49 cell line with a luciferase-expressing plasmid (Ubi-MCS-firefly_Luciferase-IRES-Puromycin) was provided by Shanghai GeneChem Co., LTD. The mice under anesthetization were injected with fLuc-MB49 cells (1 × 10⁶) into their bladder wall via a 0.33 × 12.7 mm insulin syringe (BD U-100 insulin). The D-Luciferin sodium salt (YEASEN, cat: 40901ES01) was used for the bioluminescence imaging (10 mg/kg). Three days after the cell inoculation, the bioluminescence of bladder could be detected by IVIS Lumina system (Fabris et al., 2012 (link)). The drug therapy consisted of an intravenous administration of Ag-SP-DNC (5 and 10 mg/kg) and carboplatin (10 mg/kg). A 5% glucose solution by tail-vein injection was used as the control for drug treatment. Three days post-implantation, tumor-bearing mice were randomized into groups and started drug administration once every 2 days until day 14. All animal experiments were conducted in full accordance with protocols approved by the Ethical Committee of the Qilu Hospital of Shandong University (Jinan, China).

The PZT was obtained from SCH Technology Co. Ltd. The control circuit with 660 nm µLEDs was obtained from Inper LLC. The PDMS Sylgard184 was purchased by Dow Corning. Lung cancer cell line LLC, A549, and H1299 were obtained from the Cell Bank of Chinese Academy of Sciences and cultured in Dulbecco's modified Eagle's medium (Yeasen, China) supplemented with 10% (v/v) fetal bovine serum (40 130, Yeasen, China). Chlorin e6 (Ce6) was obtained from Macklin Biochemical Technology Co., Ltd. (Shanghai, China). Imidazole ketone erastin (IKE) was purchased from Selleck Chemicals Co., Ltd. (Shanghai, China). Female BALB/c mice aged six to eight‐week were purchased from Beijing HFK Bioscience Co., Ltd. (www.hfkbio.com). Anti‐xCT (ab307601), GPX4 (ab125066), and 4‐HNE (ab46545) primary antibodies were purchased from Abcam (Shanghai, China). Anti‐FTH1 (A19544), TfR1 (A5865), ACSL4 (A6826), 4‐HNE (A24456), and β‐actin (AC026) primary antibodies were purchased from Abclonal (Wuhan, China). D‐Luciferin, sodium salt (40 902) was purchased from Yeasen (Shanghai, China).

CPT was obtained from Meryer Co., Ltd., 3,3'‐Thiodipropionic acid, 1‐ethyl‐3(3‐dimethylpropylamine) carbodiimide (EDCI) and 4‐Dimethylaminopyridine (DMAP) were purchased from Macklin Co., Ltd., Methoxypolyethylene glycol (PEG2000) and Hoechst 33 342 were obtained from Aladdin Biochemical Technology Co., Ltd. DSPE‐PEG2000 was purchased from AVT (Shanghai) Pharmaceutical Tech Co., Ltd. AZD4635 was obtained from MedChemExpress (MCE). Cy5.5 (Cyanine5.5)‐carboxylic acid was purchased from Nanjing Goyoo Biotech Co.,Ltd. 96‐well, 12‐well, 6‐well plate were obtained from NEST Biotechnology. MTT and FITC‐phalloidine were purchased from Solarbio life sciences. Annexin V‐FITC/PI Apoptosis Detection Kit and d‐Luciferin sodium salt were obtained from Yeasen Biotech Co., Ltd. Mouse Adenosine and ATP ELISA Kit were purchased from Shanghai Jianglai Biotechnology Co., Ltd. The antibodies of CRT, HMGB1 and Caspase 3 were purchased from Abcam. The antibody of GAPDH and HRP labeled sheep anti‐rabbit secondary antibody were obtained from BIOSS.
The synthesis and characterization of TST and CPT‐S‐PEG can be found in the Supporting Information.

To evaluate in vivo protein expression kinetics following transplantation of the mRNA-engineered BMSCs, bioluminescence imaging was performed using a Xenogen IVIS Lumina XRMS Series II live animal biophotonic imaging system (Caliper Life Sciences, USA). BMSCs transfected with modRNA encoding luciferase (Luc) were loaded in collagen scaffolds (n = 3) and implanted in the defect site. IVIS images were recorded at 1, 2, 3, and 4 days following surgery (n = 3 per time point). D-Luciferin, Sodium Salt (CAS 103404-75-7; Yeasen, Shanghai, China) was injected intraperitoneally using a 27 gauge needle at a dosage of 150 mg/kg body-weight under general anesthesia. After 10 min, imaging was performed with an acquiring time of 60 s, exposure time of 0.2 s, binning of 8 and field of view 24. Signals were collected from a defined region of interest (ROI) using the contour ROI tool and measured values are given in Radiance (photons/sec/cm² /steradian). Image analysis was done using the software Living Image (Caliper, USA). At 24 h time intervals following implantation, animals were sedated and non-invasively imaged. Animals with a cranial defect site and a collagen scaffold served as baseline for Luc expression. Signal intensities reported by the modRNA groups were acquired by subtracting the baseline expression.

4T1-Luc cells (1 × 10⁶) were orthotopically injected into the mammary fad of 6-week-old female BALB/c mice. Then, 25 mg/kg and 50 mg/kg HCL-23 were intraperitoneally injected (n = 4/group). The body weight and tumor size were measured every other day. The tumor volume was calculated using the formula volume (mm³) = 1/2 × a × b², where a and b are the long and short dimensions of the tumor, respectively. After D-Luciferin sodium salt (Yeasen, Shanghai, China) was injected, the IVIS Spectrum Living Imaging system (PerkinElmer, Waltham, MA, USA) was used to detect tumor bioluminescence intensity in mice. The tumor weight was recorded when the mice were sacrificed. All procedures were approved by the Animal Care and Use Committee of Guizhou Medical University (2101008).

Dulbecco’s modified eagle medium (DMEM) and fetal bovine serum (FBS) were purchased from Gibco (Thermo Fisher Scientific, USA). Trypsin-EDTA (0.25%, 1× solution), Dulbecco’s phosphate buffered saline (DPBS) and antibiotics (100 U mL⁻¹ penicillin G and 0.1 mg ml⁻¹ streptomycin) were purchased from Gibco® (Invitrogen™, Grand Island, NY). Cell Counting Kit-8 (CCK-8) was obtained from Dojindo Laboratories (Dojindo, Kumamoto, Japan). Alizarin Red S was purchased from Solarbio^®Life Sciences, Alkaline phosphatase (ALP) staining kit (BCIP/NBT Alkaline Phosphatase Color Development Kit) was purchased from Beyotime Institute of Biotechnology. Collagen was obtained from Sichuan Mingrang Tech., China and collagen cross-linking agent N-(3-(dimethylamino)-propyl)-N-Ethylcarbodiimide hydrochloride crystalline (EDC) and N-Hydroxysulfosuccinimide sodium salt (NHS) were purchased from Aladdin Industrial Corporation, Shanghai, China. Opti-MEM medium and Lipofectamine^® MessengerMAX^TM Reagent were obtained from Gibco^TM (Invitrogen, CA, USA). Human BMP-2 ELISA kit and human VEGF ELISA kit were obtained from R&D (Quantikine, R&D Systems, USA). D-Luciferin, Sodium Salt was purchased from Yeasen (Shanghai, China). All materials were utilized as received, without any further purification.