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. 2020 Oct 16;9:e57920. doi: 10.7554/eLife.57920

Figure 3. Mφ-inactivation of MT1-MMP impairs active TGFβ1 release from LAP-TGFβ1 complex.

Figure 3.

(A) Scheme of LAP-TGFβ1 complex retention in the cell surface through LAP-binding to membrane receptors. (B) Representative flow cytometry histogram plots of LAP and TGFβ1 staining in LPS-activated WT and MAC-Mmp14 KO BMDMs. (C) Standardized mean fluorescence intensity (MFI) of LAP and TGFβ1 in experiments as in B. Data are means ± SEM of seven mice per group. Unpaired t-test. (D) Western blot of transferrin receptor (TfR), α-tubulin, and LAP-TGFβ1 complex in membrane fraction (Mb), cytosolic fraction (C), and total lysate (T) from LPS-activated WT and MAC-Mmp14 KO BMDMs. (E) Quantification of LAP-TGFβ1 complex in the membrane fraction. Data are means ± SEM of 6–7 mice per genotype. Unpaired t-test. (F) Tgfb1 mRNA expression in LPS-activated WT and MAC-Mmp14 KO BMDMs. Data are means ± SEM of five mice per genotype. Unpaired t-test. (G) Arbitrary luciferase units (ALU) in HEK293 cells co-cultured with or without LPS-activated WT or MAC-Mmp14 KO BMDMs. Control corresponds to transfected HEK293 cells cultured alone. Data are means ± SEM of a representative experiment of three performed with four technical replicates per condition. One-way ANOVA followed by Tukey's multiple comparisons test. (G) ALU in HEK293 cells co-cultured with or without conditioned media from LPS-activated MAC-Mmp14 KO BMDMs transduced with mock lentivirus (GFP), or lentivirus containing full-length MT1-MMP (FL) or catalytic MT1-MMP mutant (E240A). Control corresponds to transfected HEK293 cells cultured alone. Data are means ± SEM of a representative experiment of three performed with three technical replicates per condition. One-way ANOVA followed by Tukey's multiple comparisons test.

Figure 3—source data 1. Mφ-inactivation of MT1-MMP impairs active TGFβ1 release from the LAP-TGFβ1 complex.