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. 2023 Jul 7;11:1212230. doi: 10.3389/fbioe.2023.1212230

FIGURE 3.

FIGURE 3

Myeloid cells migrate throughout all the layers of the 3D-HTLM. (A,B) Immunofluorescence of paraffin embedded and transverse sectioned 3D-HTLM. (A) Myeloid cells were detected with Abs for CD45 or CD14 (green) and nuclei were detected with DAPI. Blue and red arrows indicate myeloid cells within the top and middle layers of the model, respectively. Dotted lines outline the 3D-HTLM boundaries. Green arrow indicates the transwell membrane. (B) Abs for CD45 (red) detected myeloid cells and CD31 (green) detected HPMECs, and Sytox deep red was used to stain nuclei. A region of interest is outlined with a white box in the merged image, which is magnified in the panel to the right to highlight detail. Red and pink arrows indicate myeloid cells within the middle and bottom layers, respectively. Pink arrowheads indicate HPMEC nuclei. Green arrow indicates the transwell membrane. (C) The top SAEC layer of the 3D-HTLM was removed and probed with an Ab for CK-14 (R-PE) to locate SAECs. Nuclei were counterstained with DAPI. Images shown are representative of 40 images taken from three wells.