Figure 1.
mTurq2-Col4a1 endogenously tagged reporter design and basement membrane localization in adult mouse tissues. (A) Schematic of the COL4A1 subunit and type IV collagen trimeric protomer with an N-terminal mTurq2 fluorescent tag. (B) Schematic of the Mus musculus Col4a1 genomic locus and mTurq2 insertion site. The mTurq2 gene was inserted into Exon 2 between the coding sequences for the signal peptide and 7S domain. (C) Sagittal section of dorsal skin from mTurq2-Col4a1/+ adult mouse labeled with ColIV antibodies (magenta) and Hoechst to mark nuclei (blue; grayscale in C’ and C’’). (C’) mTurq2-COL4A1 (cyan) localizes to BM surrounding the hair follicle, where it overlaps with ColIV staining (C’’, magenta). Scale bars, 100 µm. (D–D’’) High magnification view of mTurq2-COL4A1 (cyan) localization in the interfollicular epidermis from mTurq2-Col4a1/+ adult mouse labeled with ColIV (magenta) and E-Cadherin (E-Cad; grayscale). Scale bar, 20 µm. (E–E’’) High magnification view of mTurq2-COL4A1 (cyan) localization in the interfollicular epidermis from mTurq2-Col4a1/+ adult mouse labeled with Perlecan (magenta) and E-Cadherin (E-Cad; grayscale). Scale bar, 20 µm. (F) Kidney section from mTurq2-Col4a1/+ adult mouse labeled with ColIV (magenta) and Hoechst to mark nuclei (blue). Scale bar, 100 µm. (G–G’’) High magnification view of mTurq2-COL4A1 (cyan) localization in the kidney of mTurq2-Col4a1/+ adult mouse labeled with ColIV (magenta in G; grayscale in G’) Hoechst (blue). mTurq2-COL4A1 expression can be seen in renal tubules (arrows), Bowman’s capsule (arrowheads), and the mesangial matrix (asterisks). Scale bar, 100 µm. (H) Kidney section from mTurq2-Col4a1/+ adult mouse labeled with Perlecan (magenta) and Hoechst (blue). Scale bar, 100 µm. (I–I’’) High magnification view of mTurq2-COL4A1 (cyan) localization in the kidney of mTurq2-Col4a1/+ adult mouse labeled with Perlecan (magenta in G; grayscale in G’) and Hoechst (blue). mTurq2-COL4A1 expression can be seen in renal tubules (arrows), Bowman’s capsule (arrowheads), the mesangial matrix (asterisks), and the collecting duct (CD). Scale bar, 100 µm.

mTurq2-Col4a1 endogenously tagged reporter design and basement membrane localization in adult mouse tissues. (A) Schematic of the COL4A1 subunit and type IV collagen trimeric protomer with an N-terminal mTurq2 fluorescent tag. (B) Schematic of the Mus musculus Col4a1 genomic locus and mTurq2 insertion site. The mTurq2 gene was inserted into Exon 2 between the coding sequences for the signal peptide and 7S domain. (C) Sagittal section of dorsal skin from mTurq2-Col4a1/+ adult mouse labeled with ColIV antibodies (magenta) and Hoechst to mark nuclei (blue; grayscale in C’ and C’’). (C’) mTurq2-COL4A1 (cyan) localizes to BM surrounding the hair follicle, where it overlaps with ColIV staining (C’’, magenta). Scale bars, 100 µm. (D–D’’) High magnification view of mTurq2-COL4A1 (cyan) localization in the interfollicular epidermis from mTurq2-Col4a1/+ adult mouse labeled with ColIV (magenta) and E-Cadherin (E-Cad; grayscale). Scale bar, 20 µm. (E–E’’) High magnification view of mTurq2-COL4A1 (cyan) localization in the interfollicular epidermis from mTurq2-Col4a1/+ adult mouse labeled with Perlecan (magenta) and E-Cadherin (E-Cad; grayscale). Scale bar, 20 µm. (F) Kidney section from mTurq2-Col4a1/+ adult mouse labeled with ColIV (magenta) and Hoechst to mark nuclei (blue). Scale bar, 100 µm. (G–G’’) High magnification view of mTurq2-COL4A1 (cyan) localization in the kidney of mTurq2-Col4a1/+ adult mouse labeled with ColIV (magenta in G; grayscale in G’) Hoechst (blue). mTurq2-COL4A1 expression can be seen in renal tubules (arrows), Bowman’s capsule (arrowheads), and the mesangial matrix (asterisks). Scale bar, 100 µm. (H) Kidney section from mTurq2-Col4a1/+ adult mouse labeled with Perlecan (magenta) and Hoechst (blue). Scale bar, 100 µm. (I–I’’) High magnification view of mTurq2-COL4A1 (cyan) localization in the kidney of mTurq2-Col4a1/+ adult mouse labeled with Perlecan (magenta in G; grayscale in G’) and Hoechst (blue). mTurq2-COL4A1 expression can be seen in renal tubules (arrows), Bowman’s capsule (arrowheads), the mesangial matrix (asterisks), and the collecting duct (CD). Scale bar, 100 µm.

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