Skip to main content
. 2023 Jul 20;12(14):1899. doi: 10.3390/cells12141899

Figure 2.

Figure 2

Validation of the interaction between TIMP-1 and CD74 by immunocytochemistry and immunoprecipitation analysis. (a) Colocalization of TIMP-1 and CD74 in THP-1 monocytes. Fluorescence microscopy analysis costained with anti-TIMP-1/Star635 secondary antibody (red), anti-CD74/Star488 secondary antibody (green), and DAPI (blue, DNA labeling in nuclei). The merged image shows the overlay (yellow) of the two channels demonstrating TIMP-1 colocalization with CD74. Arrows indicate membrane regions of substantial colocalization. Scale bar: 10 µm. (b,c) Immunoprecipitation of TIMP-1 with cell surface CD74. Semi-endogenous pull-down assay using protein cell lysate from HEK29 cells transfected with FLAG-tagged CD74 and recombinant TIMP-1 analyzed by Western blotting with anti-CD74 (b) and anti-TIMP-1 (c) antibodies. (d,e) TIMP-1 interacts with the soluble CD74 ectodomain fragment (sCD74). Immunoprecipitation of His-tagged sCD74 and recombinant TIMP-1 and the analysis of precipitated proteins by Western blotting using anti-His (d) and anti-TIMP-1 (e) antibodies. C: control.