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. 2024 Nov 9;27(12):111355. doi: 10.1016/j.isci.2024.111355

Figure 1.

Figure 1

Sept-7 localizes in spatially and temporally distinct patterns in FA sub-populations

(A) Representative TIRFM images of MEFs stained with paxillin (magenta), F-actin (cyan), and Sept-7, (yellow, low, and high contrast) of cells plated on 10 μg/mL FN. Magenta and cyan boxes highlight perinuclear and peripheral FAs respectively; scale bar, 30 μm.

(B) Insets of perinuclear (magenta), and peripheral (cyan) FAs from (A); scale bar, 5 μm.

(C) Curve plots showing normalized line scan intensities of paxillin (magenta), Sept-7 (yellow), and F-actin (cyan), for perinuclear (left) and peripheral (right) FAs, shaded curves represent SD, n = 3 replicates, 38–43 line scans.

(D and E) Quantification of colocalization between paxillin and Sept-7, and F-actin and Sept-7 respectively, for perinuclear and peripheral FAs, n = 3 replicates, 1050–2270 FAs.

(F and G) Montages taken from live TIRFM movie of perinuclear and peripheral FAs from MEFs co-transfected with paxillin-mCherry (gray, top) and Sept-7-YFP (fire, middle), and merged channels (bottom). Selected FAs are circled, and colocalized Sept-7 indicated with yellow arrows; scale bar, 5 μm.

(H) Quantification of the minimum Sept-7-paxillin localized lifetime, n = 3 replicates, 32–40 FAs. Statistical analysis was performed using Mann-Whitney test, ∗∗∗∗p < 0.0001, orange horizonal lines show medians.