Journal: The Journal of Biological Chemistry
Article Title: Loss of integrin alpha7-mediated signaling induces a dendritic cell-like phenotype in macrophages cultured on laminin-211/221 isoforms
doi: 10.1016/j.jbc.2025.110419
Figure Lengend Snippet: Localization of laminin-related integrins and the effect of functional blocking of integrin α7 in THP-1 macrophages cultured on laminin-211/221/221E8 or laminin-511 for 5 days . A , morphology and multiple immunofluorescent staining for laminin-related integrins (α7, α6, and α3) and phalloidin in THP-1 macrophages cultured on laminin (LM)-211/-511. Nuclei were counterstained with DAPI. Immunostaining was performed four times independently, imaging a total of 300 to 350 cells per antibody. B , morphology of THP-1 macrophages cultured on laminin-211/-221/-221E8 in the presence of integrin α7 function-blocking antibodies (anti-α7) or isotype control IgG. In the presence of integrin α7 function-blocking antibodies, dendritic-like branching and extended cellular processes are observed. These experiments were repeated at least five times independently. C , morphology of THP-1 macrophages cultured on laminin-511 in the presence of integrin α7 function-blocking antibodies. D , western blotting of integrin α7 under conditions of integrin α7-targeted small interfering RNA (siITGA7) on day 5. siITGA7 #9 and #1 reduced integrin α7 levels on laminin-211. Nonspecific siRNA was used as control (siCtrl); β-actin was used as a loading control. E , gene expression levels of integrin α7. Data are presented as the mean ± SEM of three independent experiments. ∗ p < 0.05, ∗∗∗ p < 0.001 and ∗∗∗∗ p < 0.0001 as determined by Tukey’s post hoc test. F , morphology of THP-1 macrophages treated with integrin α7-targeted small interfering RNA #9, #1 or nonspecific siRNA on day 5. The knockdown experiment was repeated three times independently. Integrin α7 knockdown induced dendritic processes in THP-1 macrophages cultured on laminin-211 ( arrows ). Nonspecific siRNA did not induce obvious morphological alterations. The scale bars represent 10 μm. DAPI, 4′,6-diamidino-2-phenylindole; IgG, immunoglobulin G.
Article Snippet: An isotype control IgG was obtained from Bio X Cell (BE0366).
Techniques: Functional Assay, Blocking Assay, Cell Culture, Staining, Immunostaining, Imaging, Control, Western Blot, Small Interfering RNA, Gene Expression, Knockdown
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