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. 2023 May 6;24(9):8365. doi: 10.3390/ijms24098365

Figure 3.

Figure 3

Functional consequences of TNC CCL2 interaction on migration in vitro: (A) CCR2 expression of murine bone marrow cells stimulated with M-CSF or GM-CSF for 7 days on mRNA expression level relative to the housekeeping gene HPRT, n = 3, mean +/− SEM. (B) CCL2 protein amount determined via ELISA in supernatant of murine bone marrow cells stimulated with GM-CSF for 7 days, n = 5, mean +/− SEM. (C) Schematic depiction of transwell migration assay of mBMMs (added to the upper chamber) towards a gradient of mCCL2 (added to the lower chamber). The bottom of the insert was coated with matrix molecules. mBMMs that transmigrated through the membrane and adhered to the bottom of the insert were quantified. (D) Transwell migration assay with mBMMs and coating of the bottom surface of the insert with BSA or TNC or FN without chemoattractant or towards 20 nM CCL2. Quantification of cells on matrix coating after 3 h of migration by counting DAPI stained nuclei. Average number of cells/field is shown, n = 3, mean +/− SEM. (E) Chemotactic index calculated of 3D. Student’s t test was used to compare datasets. ** p < 0.01, * p < 0.05, ns ≥ 0.05. (F) mBMMs seeded on PBS, TNC or FN and imaged after 1 h, 40× magnification, n = 3, representative images are shown. Scale bar is 100 µm.