Figure 4. hSNX16 CC mutants exhibit... | Rockefeller University Press

Figure 4.

$Figure 4. hSNX16 CC mutants exhibit altered subcellular localization in vivo. hSNX16ΔCC reduces and hSNX163A enhances hSNX16 punctate localization in mammalian cells. (A) Representative images of cell body and neurites from immunostained DIV 7 rat hippocampal neurons transiently expressing the indicated myc-hSNX16 variants. (B) Representative images of the indicated cell lines expressing indicated hSNX16 CC variants and fixed 24 h after transfection. hSNX16-transfected U2OS cells were stained with α-EEA1 antibodies. (C) CoV (standard deviation/mean of pixel intensities) of myc-hSNX16 CC mutants in hippocampal cell bodies. Quantification is from ≥35 neurons per condition from three independent coverslips, tested for normality and analyzed using a one-way ANOVA followed by Tukey’s test. (D) Histograms depict fraction of pixels at the indicated intensities for hSNX16 and hSNX163A in HeLa cells. Quantification is from 14 cells per condition and analyzed using a Mann–Whitney U test within each bin. All images show 2D maximum intensity projections of confocal stacks. Data are shown as box-and-whisker plots with all data points superimposed in C and as mean + SEM in D. *, P < 0.05; ***, P < 0.001. ns, not significant. Scale bars, 10 µm.$

hSNX16 CC mutants exhibit altered subcellular localization in vivo. hSNX16^ΔCC reduces and hSNX16^3A enhances hSNX16 punctate localization in mammalian cells. (A) Representative images of cell body and neurites from immunostained DIV 7 rat hippocampal neurons transiently expressing the indicated myc-hSNX16 variants. (B) Representative images of the indicated cell lines expressing indicated hSNX16 CC variants and fixed 24 h after transfection. hSNX16-transfected U2OS cells were stained with α-EEA1 antibodies. (C) CoV (standard deviation/mean of pixel intensities) of myc-hSNX16 CC mutants in hippocampal cell bodies. Quantification is from ≥35 neurons per condition from three independent coverslips, tested for normality and analyzed using a one-way ANOVA followed by Tukey’s test. (D) Histograms depict fraction of pixels at the indicated intensities for hSNX16 and hSNX16^3A in HeLa cells. Quantification is from 14 cells per condition and analyzed using a Mann–Whitney U test within each bin. All images show 2D maximum intensity projections of confocal stacks. Data are shown as box-and-whisker plots with all data points superimposed in C and as mean + SEM in D. *, P < 0.05; ***, P < 0.001. ns, not significant. Scale bars, 10 µm.