Fig. 1 | Nature Communications

Fig. 1

From: Spatial cycles mediated by UNC119 solubilisation maintain Src family kinases plasma membrane localisation

Fig. 1

Steady-state PM localisation of Src and Fyn requires vesicular traffic from the RE and Golgi. a Confocal images showing the steady-state localisation of mCitrine fused Src and Fyn in HeLa cells co-expressing the PM marker mCh-tK-Ras, and stained with antibodies against the RE compartment marker, Rab11a (see also Supplementary Fig. 2a). Dot plot depicts Pearson’s correlation coefficient for Src and Fyn with either the PM marker or the RE marker (n > 60 cells per condition from two independent experiments, data are mean ± SD). b HeLa cells co-expressing the dominant-negative mutant form of Rab11 (Rab11S25N-BFP) and either Src-mCit or Fyn-mCit (see also Supplementary Fig. 2b). Dot plot depicts the percentage of the area of Src or Fyn pixels staining internal membrane structures that overlapped with the Rab11S25N-BFP pixels (n > 40 cells per condition, data are mean ± SD, ****P < 0.0001, Student’s t-test). c Confocal micrographs of HeLa cells expressing Src-mCit and Fyn-mCit, transfected with Rab11a/b targeting siRNA or non-targeting siRNA control nucleotides. d Representative western blot showing Rab11a levels of transfected cells and the Tubulin-loading control (n = 3 independent experiments). e HeLa cells co-expressing the Golgi marker (GalT-mCer) and Src-mCit or Fyn-mCit, either cultured at 37 °C (left panels) or 20 °C (right panels) for 24 h, at which point cells were fixed. Graph depicts the percentage of the area of Src or Fyn pixels staining internal membrane structures that overlapped with the GalT-mCer pixels (n > 40 cells for each condition from two independent experiments, data are depicted as a box and whiskers plot, showing the median and the full range of the measurements. ***P < 0.001; ****P < 0.0001; n.s., not significant, as determined by Student’s t-test). f Time series of HeLa cells co-expressing the Golgi marker (GalT-mCer) and Fyn-mCit, treated with the 25 μM of the APT inhibitor palmostatin-M (Pal). The plot to the right depict the integrated intensity of Fyn-mCit at the Golgi over the integrated intensity over the whole of the cell (n = 17 cells from two independent experiments; data are mean ± SEM). Scale bars, 10 μm

Back to article page