Supporting Information. Light-enhanced hypoxia-response of conjugated polymer nanocarrier. for successive synergistic photodynamic and chemo-therapy

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1 Supporting Information Light-enhanced hypoxia-response of conjugated polymer nanocarrier for successive synergistic photodynamic and chemo-therapy Xiaolong Zhang a,d, Ming Wu a,d, Jiong Li a,c,d, Shanyou Lan a,b, Yongyi Zeng a,b,d, Xiaolong Liu a,d*, and Jingfeng Liu a,b,d* a. The United Innovation of Mengchao Hepatobiliary Technology Key Laboratory of Fujian Province, Mengchao Hepatobiliary Hospital of Fujian Medical University, Fuzhou , P. R. China b. Liver Disease Center, The First Affiliated Hospital of Fujian Medical University, Fuzhou , P. R. China c. School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou , P.R. China d. The Liver Center of Fujian Province, Fujian Medical University, Fuzhou , P. R. China S-1

2 Figure S1. GPC results of the CPs. (A) GPC elution curves of CPs. (B) Summary of molecular weight and polydispersity of CPs Figure S2. FT-IR spectra of 4,4 -azodianiline, terephthalaldehyde and CPs. S-2

3 Figure S3. The chemical structure of CPT, Ce6 and PVP. Figure S4. Zeta potential of CPs-CPT-Ce6 NPs. S-3

4 Figure S5. Stability of the CPs-CPT-Ce6 in different solution for different times. Photographs of CPs-CPT-Ce6 NPs in different solution for 0 day (A) and 5 days (B). (C) Average size changes of CPs-CPT-Ce6 NPs in different solution for 5 days. Error bars are based on triplicate measurements. Figure S6. Fluorescence emission spectra (A) and standard curve with linear calibration (B) of CPT at different concentrations. Error bars are based on triplicate measurements. The error bars represent standard errors (n=3). S-4

5 Figure S7. The absorbance of 9,10-dimethylanthracene (ABDA, 50 µm) after photodecomposition by ROS generation with (A) or without (B) 670 nm laser irradiation at 50 mw/cm 2 in the presence of CPs-CPT-Ce6 NPs. Figure S8. The cumulative CPT release kinetics of CPs-CPT-Ce6 NPs with or without Na 2 S 2 O 4 treatment in PBS buffer at 37 C. Error bars are based on triplicate measurements. The error bars represent standard errors (n=3). S-5

6 Figure S9. Cell viabilities of HeLa cells and NIH3T3 cells treated with different concentration of CPs NPs for 24 h without irradiation. The error bars represent standard errors (n=3). Figure S10. Cell viability of HeLa cells treated with different concentration of CPs-CPT-Ce6 NPs and then with or without laser irradiation under normoxia condition. The error bars represent standard errors (n=3). The statistical analysis was performed with two-tailed paired Student s T-test (*p < 0.05, **p < 0.01, ***p < 0.001). S-6

7 Figure S11. The cell apoptosis and necrosis were analyzed by flow cytometry analysis using Annexin V-FITC and PI staining when the HeLa cells treated with 20 µg/ml of CPs NPs, CPs-CPT NPs, CPs-Ce6 NPs and CPs-CPT-Ce6 NPs and then with or without laser irradiation under normoxia condition. Figure S12. Confocal images of HeLa cells that incubated with CPs-CPT-Ce6 NPs under hypoxia and normoxia conditions. Scale bar is 20 µm. S-7

8 Figure S13. DIC images of HeLa cells treated with CPs NPs, CPs-CPT NPs, CPs-Ce6 NPs and CPs-CPT-Ce6 NPs and then with laser irradiation under PDT induced hypoxic environment. The morphology of dead cells became shrinkage and round. Scale bar is 100 µm. S-8