Figure 4. Spatial distribution and... | Rockefeller University Press

Figure 4.

$Spatial distribution and correlation between key basement membrane components in mTurq2-Col4a1/+ embryonic backskins resembles that of WT littermates. (A) Average intensity projection (XZ optical reconstruction) of backskin from WT E15.5 embryo labeled with Hoechst (merge, top), anti-laminin β-1 antibody (A’), and anti-collagen IV antibody (A’’). No signal is observed in the mTurq2 channel (A’’’). (B) As in A, except backskin from a mTurq2-Col4a1/+ embryo. Robust mTurq2-COL4A1 signal recapitulates the labeling pattern of the collagen IV antibody. (C) Pearson’s correlation coefficient (r) measuring the correlation between laminin β-1 and collagen IV expression (antibody labeling) in WT and mTurq2-Col4a1/+ E15.5 backskins as labeled. Larger datapoints—biological replicate means (n = 3 embryos), smaller datapoints—individual images per replicate (n = 10 per embryo). (D) Fraction of laminin β-1 contribution to the laminin β-1:collagen IV overlap in E15.5 backskins; refer to the main text for methodology. (E) As in D, except collagen IV contribution to laminin β-1:collagen IV overlap. Dots represent individual images (n = 10 per embryo) and boxes denote the minimum and maximum mean per biological replicate (n = 3 embryos). (F) Average intensity projection (XZ optical reconstruction) of backskin from WT E15.5 embryo labeled with Hoechst (merge, top), anti-laminin β-1 antibody (F’), and anti-perlecan antibody (F’’). (G) As in F, except backskin from an mTurq2-Col4a1/+ embryo. (H–J) As in C–E, except measuring the correlation and overlap fractions between laminin β-1 and perlecan. (K–T) As in A–J, except backskins are from E12.5 embryos. Scale bars, 20 µm. ns = not significant, Mann–Whitney U test.$

Spatial distribution and correlation between key basement membrane components in mTurq2-Col4a1/+ embryonic backskins resembles that of WT littermates. (A) Average intensity projection (XZ optical reconstruction) of backskin from WT E15.5 embryo labeled with Hoechst (merge, top), anti-laminin β-1 antibody (A’), and anti-collagen IV antibody (A’’). No signal is observed in the mTurq2 channel (A’’’). (B) As in A, except backskin from a mTurq2-Col4a1/+ embryo. Robust mTurq2-COL4A1 signal recapitulates the labeling pattern of the collagen IV antibody. (C) Pearson’s correlation coefficient (r) measuring the correlation between laminin β-1 and collagen IV expression (antibody labeling) in WT and mTurq2-Col4a1/+ E15.5 backskins as labeled. Larger datapoints—biological replicate means (n = 3 embryos), smaller datapoints—individual images per replicate (n = 10 per embryo). (D) Fraction of laminin β-1 contribution to the laminin β-1:collagen IV overlap in E15.5 backskins; refer to the main text for methodology. (E) As in D, except collagen IV contribution to laminin β-1:collagen IV overlap. Dots represent individual images (n = 10 per embryo) and boxes denote the minimum and maximum mean per biological replicate (n = 3 embryos). (F) Average intensity projection (XZ optical reconstruction) of backskin from WT E15.5 embryo labeled with Hoechst (merge, top), anti-laminin β-1 antibody (F’), and anti-perlecan antibody (F’’). (G) As in F, except backskin from an mTurq2-Col4a1/+ embryo. (H–J) As in C–E, except measuring the correlation and overlap fractions between laminin β-1 and perlecan. (K–T) As in A–J, except backskins are from E12.5 embryos. Scale bars, 20 µm. ns = not significant, Mann–Whitney U test.