The non-classical nuclear import carrier Transportin 1 modulates circadian rhythms through its effect on PER1 nuclear localization
Fig 2
Knockdown and knockout of TNPO1 shortens the circadian period.
(A) Left: Representative time series for the effect of three different shTnpo1 constructs compared to a non-silencing (ns) control shRNA. Right: Mean period deviation and relative Tnpo1 mRNA and protein levels. Bottom: representative western blot of TNPO1 protein levels in ns control and Tnpo1 depleted cells; lanes are from the same blot with identical exposure time. Error bars = SD, nperiod deviation = 7 to 14 individual recordings, nmRNA and protein = 4–10 individual cell lysates; one-sample t-test to test whether period deviations are different from zero, *** p < 0.001. (B) Top: Schematic illustration of the CRISPR/Cas9-mediated genome editing of the Tnpo1 gene. Bottom left: Representative time series of U-2 OS reporter cells transduced with Cas9 expression vector and indicated guide RNA (gRNA). Middle: Mean periods of independent bioluminescence recordings (error bars = SD, n = 3, one-way ANOVA with Bonferroni posthoc test comparing sgRNAs to wild-type: *** p < 0.001, n.s. = non- significant. Right: Representative western blot of residual TNPO1 protein levels in genome-edited cells.