Fig 2: The 10 Hz rTMS inhibited OGD/R injury induced M1 polarization of microglia. Immunostaining for CD206 (a) and iNOS (b) in BV2 cells with indicated treatment. c, d Western blots and quantification of CD206 and iNOS in each group. n = 3. Data are expressed as mean ± SD; *P < 0.05, **P < 0.01, ***P < 0.001 versus Ctrl group; ##P < 0.01, ###P < 0.001 versus OGD/R group. Scale bar: 10 µm

Fig 3: Interaction between BMECs and HCC cells enhances tumor growth in the spine. (A) si-CX3CL1 was transfected into BMECs to construct CX3CL1low BMECs. Transwell assays were used to examine recruitment of BMECs co-cultured with or without HMCC97H cells. The number of the recruited cells was quantified. n=3. Cell recruitment were analyzed using ANOVA test. (B) Cell vitality of BMECs was assessed using CCK-8 assay; n=3. Data were analyzed using ANOVA followed by Tukey's test. (C) Tumor size was measured based on the volume. n=5. Scale bar, 5 mm. Data were analyzed using an unpaired t-test. (D) Samples from 2 groups were examined using immunofluorescence. Magnification, ×20. The result was analyzed using an unpaired t-test. Gray bars represent the CX3CL1nor BMECs group and black bars represent the CX3CL1low BMECs group (E) Flow cytometry revealed the percentage and number of F4/80+CD11b+ cells; n=5. (F) Flow cytometry was used for detecting expression of M2 macrophage surface markers, CX3CR1, MRC1 and CD163; n=5. Cells were selected based on higher expression of CD45 for macrophages. Subsequently, CD11b and F4/80 expression were used for further characterization of macrophages. FACS (fluorescent-activated cell sorting) data were analyzed using a Kruskal Wallis test followed by Dunn's non-parametric post hoc test. HCC, hepatocellular carcinoma; BMECs, bone marrow endothelial cells; si, small interfering; CCK-8, Cell Counting kit-8; CX3CL1, C-X3-C motif chemokine ligand 1.

Fig 4: CX3CL1 promotes the invasion and migration of HCC cells through Src. A total of 50 or 100 nM recombinant human CX3CL1 was added to the medium. CX3CL1 antibody was added to neutralize free CX3CL1 in medium. Bosutinib was used to inhibit Src. (A) Representative images and statistical analysis of Transwell migration assays showed the migratory ability of HCC cells. The number of cells which had invaded was quantified; n=5; scale bar, 200 µm. (B) Representative images and bar graphs of Transwell invasion assays showed the invasive ability of HCC cells after 24 h. The number of cells which had migrated was quantified; n=5; scale bar, 200 µm. Cell migration and invasion were analyzed using ANOVA. (C) Expression of THP-1 cells surface markers MRC1 and CD163 were detected by western blot analysis; n=3. Western blot analysis data were analyzed using a Student's t-test following log transformation. HCC, hepatocellular carcinoma; CX3CL1, C-X3-C motif chemokine ligand 1; MRC1, mannose receptor C-type 1.

Fig 5: Investigation of the biocompatibility and needle insertion of the biodegradable microneedle sensor through staining and imaging.(A) H&E staining (right) of the mouse skin after microneedle insertion (at day 0, day 3, and day 10; scale bar, 200 µm). (B) Immunofluorescence from 4',6-diamidino-2-phenylindole (DAPI), CD206, and CD68 staining of the mouse skin after device-insertion surgery detected at day 1, day 7, and day 14. The green and red fluorescence are macrophages: green (M2), red (total macrophages). (C) Investigation of the biodistribution in the main urinary organs using an in vivo imaging system (IVIS) after intraperitoneal injection of the glucose-responsive fluorescent monomer (GF-monomer) solution.