Additional file 1 of Site-specific nanomodulator capable of modulation apoptosis for enhanced colorectal cancer chemo-photothermal therapy
Creators: name: Wang, Shuqi
name: Zhou, Li
name: Tian, Hailong
name: Li, Bowen
name: Su, Miao
name: Li, Qiong
name: Nice, Edouard C.
name: Huang, Canhua
name: Shao, Jichun
name: He, Tao
Publisher: name: figshare
Year: 2023
Subjects: Biochemistry
Space Science
Medicine
Cell Biology
Molecular Biology
Pharmacology
Biotechnology
Chemical Sciences not elsewhere classified
Biological Sciences not elsewhere classified
Cancer

description: Additional file 1: Figure S1. Preparation and Characterization of CLT-IR820 and stability of HCR NPs. (A) Schematic illustration of the synthesis of HA-HCQ. (B) 1H NMR results of HA-HCQ. (C) The Tyndall effect of HCR NPs at the indicated time points. (D) The Tyndall effect of CLT-IR820. (E) Size distribution of CLT-IR820. (F) UV-vis absorption spectra of CLT-IR820. (G) Zeta potential of CLT-IR820. (H) TEM image of CLT-IR820. Scale bar: 100 nm. Figure S2. Cytotoxicity of HCR NPs in vitro. (A-B) Quantification of DLD-1 and HCT116 cells co-cultured with CLT, CLT+IR820+Laser, CLT-IR820+Laser, and HCR NPs+Laser in the EDU assay. (λ=808 nm, P =1.0 W/cm2; 40 s). ***P &lt; 0.001. (C-E) Representative images of the colony formation of DLD-1 and HCT116 cells treated with HA-HCQ, CLT, IR820+Laser, CLT+ IR820+Laser, CLT- IR820+Laser, and HCR NPs+Laser. (λ=808 nm, P =1 W/cm2; 60 s). ***P &lt; 0.001. (F) Cell imaging showing the survival of DLD-1 and HCT116 cells treated with CLT, CLT+ IR820+Laser, CLT- IR820+Laser, and HCR NPs+Laser. Live cells were marked with Calcein-AM (green fluorescence), while dead cells were marked with propidium iodide (red fluorescence). Scale bar: 50 µm. (λ=808 nm, P =1 W/cm2; 2 min). Figure S3. Apoptosis and inhibition of autophagy. (A) Fluorescent images of JC-1-stained DLD-1 and HCT116 cells, including CLT, CLT+IR820+Laser, CLT-IR820+Laser, and HCR NPs+Laser treatments. Scale bar: 50 μm. (λ=808 nm, P=1 W/cm2; 2 min). (B) Fluorescent images of DQBSA-stained DLD-1 and HCT116 cells, including HA-CLT-IR820, and HCR NPs treatments. Scale bar: 50 μm. (λ=808 nm, P=1 W/cm2; 2 min). (C) Immunoblot analysis of p62 and LC3-II in DLD-1 and HCT116 cells treated with CLT, HA-CLT-IR820, and HCR NPs. (λ=808 nm, P=1 W/cm2; 2 min). Figure S4. HCR NPs-mediated ROS inhibited colorectal cancer cell survival. (A-B) Flow cytometry statistical analysis of intracellular ROS generation in DLD-1 and HCT116 cells treated with HCR NPs+Laser with or without NAC treatment using DCFH-DA as a probe. (λ=808 nm, P=1 W/cm2; 2 min). (C-E) The proliferation of DLD-1 and HCT116 cells treated with HCR NPs+Laser with or without NAC treatment as measured by EdU assay. Scale bar: 50 μm. (λ=808 nm, P =1 W/cm2; 40 s). ***P &lt; 0.001. (F-G) Flow cytometry analysis of intracellular ROS generation in DLD-1 and HCT116 cells treated with IR820+Laser or HCR NPs+Laser using DCFH-DA as a probe. (λ=808 nm, P=1 W/cm2; 2 min). Figure S5. The photothermal capacity of HCR NPs in vivo. (A) Schematic illustration of the photothermal capacity of HCR NPs in vivo. (B) Fluorescence images of tumors and major organs at 24 h after injection with IR820, CLT-IR820, free HA+HCR NPs, and HCR NPs. (C) In vivo thermal analysis of different groups of treated mice. (λ=808 nm, P=1 W/cm2; 5 min).