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. 2023 Apr 9;13(7):2226–2240. doi: 10.7150/thno.81325

Figure 4.

Figure 4

Ropivacaine-doped hydrogels increase MHC-I in tumor cells. (A) Schematic of MHC-I upregulation by ropivacaine-loaded hydrogels. AP: autophagosome, AL: autolysosome, Rop: ropivacaine. (B) Fluorescent image of LC3 and lysosomes. Ad-GFP-LC3B-infected 4T1 cells were treated with PF, PFR, PFI, or PFIR for 8 h and then stained with Lyso-Tracker Red. Scale bar is 5 μm. (C) Statistical results of the size of lysosomes from (B). (D-F) Western blot and statistical results of LC3II and P62. Cells were subjected to different treatments for 8 h. (G) Flow cytometry results of the mean fluorescence intensity (MFI) of MHC-I in 4T1 cell. (H) Statistical results from (G). (I) Immunofluorescent staining of MHC-I (red), LAMP2 (green), and DAPI (blue). Scale bar is 5 μm. All the experiments were repeated three times. Data represents mean ± SD. *P < 0.05, **P < 0.01 and ***P < 0.001. 'NS' to indicate non-significance. PF: PF127 hydrogel, PFR: ropivacaine-loaded PF127 hydrogel, PFI: ICG-loaded PF127 hydrogel, PFIR: ropivacaine and ICG co-loaded PF127 hydrogel.