bioimagesUnknownPatricia L. YeyatiThe Journal of Cell Biologyhttps://www.ebi.ac.uk/biostudies/studies/S-JCBD-201607032JCB

RPE1 WT cells maintained in the presence of CK-869 for three hours while ciliogenesis is promoted through serum depletion.

RPE1 WT cells mantained in the presence of CK-869 (10 uM) for three hours while ciliogenesis is promoted through serum depletion

Effect of Cytochalasin D treatment (4 hours 2 microMolar in FCS) on the endogenous distribution of IFT81. Cells were fixed in methanol and stained with antibosies for IFT81, gamma and acetylated tubulin.

RPE1 WT cells mantained in the presence of CK-869 for three hours while ciliogenesis is promoted through serum depletion

RPE1 WT cells mantained in the presence of CK-869 (10 uM) for three hours while ciliogenesis is promoted through serum depletion.

KDM3Anull cells maintained in the presence of CK-869 for three hours while ciliogenesis is promoted through serum depletion.

KDM3Anull cells maintained in the presence of CK-869 for three hours while ciliogenesis is promoted through serum depletion.

Cells were serum depleted for 3 hours, untreated or treated with the indicated concentrations of CK-869 or DMSO as solvent control .

Effect of acute ARP2/3 inhibition (3 hours) on the endogenous distribution of IFT81 when RPE1 WT cells are treated with 50-100 uM CK-869. Cells were fixed in methanol and stained with antibodies to IFT81 , gamma and acetylated tubulin.

Distribution of endogenous IFT81 in untreated wild type RPE1 cultures (3 hours in low serum in solvent controls).

Distribution of IFT81 in untreated (solvent control) RPE1 WT cells mantained in FCS

bioimagesImage 641825 (Figure 6G: Effect of Cytochalasin D on IFT of RPE1 WT cells - None)Image 641841 (Figure 6G: Differential sensitivity between RPE1 and KDM3A null cilia to CK-869 - RPE1 CK-869)Image 641811 (Figure 6G: Effect of ARP2/3 inhibition on IFT in RPE1 WT cells - None)Image 641810 (Figure 6G: Effect of ARP2/3 inhibition on IFT in RPE1 WT cells - None)Figure 6G: Effect of Cytochalasin D on IFT of RPE1 WT cells - NoneImage 641829 (Figure 6G: Effect of Cytochalasin D on IFT of RPE1 WT cells - None)Image 641846 (Figure 6G: Differential sensitivity between RPE1 and KDM3A null cilia to CK-869 - KDM3Anull CK-869)Image 641838 (Figure 6G: Differential sensitivity between RPE1 and KDM3A null cilia to CK-869 - RPE1 CK-869)Figure 6G: Differential sensitivity between RPE1 and KDM3A null cilia to CK-869 - RPE1 CK-869Image 641821 (Figure 6G: Effect of Cytochalasin D on IFT of RPE1 WT cells - None)Image 641827 (Figure 6G: Effect of Cytochalasin D on IFT of RPE1 WT cells - None)Image 641808 (Figure 6G: Effect of ARP2/3 inhibition on IFT in RPE1 WT cells - None)Image 641823 (Figure 6G: Effect of Cytochalasin D on IFT of RPE1 WT cells - None)Image 641812 (Figure 6G: Effect of ARP2/3 inhibition on IFT in RPE1 WT cells - None)Figure 6G: Effect of Cytochalasin D on IFT of RPE1 WT cellsImage 641809 (Figure 6G: Effect of ARP2/3 inhibition on IFT in RPE1 WT cells - None)Image 641840 (Figure 6G: Differential sensitivity between RPE1 and KDM3A null cilia to CK-869 - RPE1 CK-869)Image 641839 (Figure 6G: Differential sensitivity between RPE1 and KDM3A null cilia to CK-869 - RPE1 CK-869)Image 641843 (Figure 6G: Differential sensitivity between RPE1 and KDM3A null cilia to CK-869 - KDM3Anull CK-869)Image 641813 (Figure 6G: Effect of ARP2/3 inhibition on IFT in RPE1 WT cells - None)Image 641845 (Figure 6G: Differential sensitivity between RPE1 and KDM3A null cilia to CK-869 - KDM3Anull CK-869)Image 641832 (Figure 6G: Effect of ARP2/3 inhibition on IFT in RPE1 WT cells - RPE1 Wild Type Cell)Image 641814 (Figure 6G: Effect of ARP2/3 inhibition on IFT in RPE1 WT cells - None)Image 641815 (Figure 6G: Effect of ARP2/3 inhibition on IFT in RPE1 WT cells - None)Figure 6G: Differential sensitivity between RPE1 and KDM3A null cilia to CK-869 - KDM3Anull CK-869Image 641836 (Figure 6G: Differential sensitivity between RPE1 and KDM3A null cilia to CK-869 - RPE1 CK-869)Image 641834 (Figure 6G: Effect of ARP2/3 inhibition on IFT in RPE1 WT cells - RPE1 Wild Type Cell)Image 641807 (Figure 6G: Effect of ARP2/3 inhibition on IFT in RPE1 WT cells - None)Image 641819 (Figure 6G: Effect of Cytochalasin D on IFT of RPE1 WT cells - None)Image 641816 (Figure 6G: Effect of ARP2/3 inhibition on IFT in RPE1 WT cells - None)Image 641835 (Figure 6G: Differential sensitivity between RPE1 and KDM3A null cilia to CK-869 - RPE1 CK-869)Image 641820 (Figure 6G: Effect of Cytochalasin D on IFT of RPE1 WT cells - None)Image 641817 (Figure 6G: Effect of ARP2/3 inhibition on IFT in RPE1 WT cells - None)Image 641822 (Figure 6G: Effect of Cytochalasin D on IFT of RPE1 WT cells - None)Image 641828 (Figure 6G: Effect of Cytochalasin D on IFT of RPE1 WT cells - None)Image 641826 (Figure 6G: Effect of Cytochalasin D on IFT of RPE1 WT cells - None)Image 641824 (Figure 6G: Effect of Cytochalasin D on IFT of RPE1 WT cells - None)Image 641848 (Figure 6G: Differential sensitivity between RPE1 and KDM3A null cilia to CK-869 - KDM3Anull CK-869)Figure 6G: Effect of ARP2/3 inhibition on IFT in RPE1 WT cellsImage 641830 (Figure 6G: Effect of Cytochalasin D on IFT of RPE1 WT cells - None)Image 641818 (Figure 6G: Effect of ARP2/3 inhibition on IFT in RPE1 WT cells - None)Figure 6G: Effect of ARP2/3 inhibition on IFT in RPE1 WT cells - NoneImage 641837 (Figure 6G: Differential sensitivity between RPE1 and KDM3A null cilia to CK-869 - RPE1 CK-869)Figure 6G: Effect of ARP2/3 inhibition on IFT in RPE1 WT cells - RPE1 Wild Type CellImage 641831 (Figure 6G: Effect of ARP2/3 inhibition on IFT in RPE1 WT cells - RPE1 Wild Type Cell)Image 641842 (Figure 6G: Differential sensitivity between RPE1 and KDM3A null cilia to CK-869 - RPE1 CK-869)Figure 6G: Differential sensitivity between RPE1 and KDM3A null cilia to CK-869Image 641844 (Figure 6G: Differential sensitivity between RPE1 and KDM3A null cilia to CK-869 - KDM3Anull CK-869)Image 641847 (Figure 6G: Differential sensitivity between RPE1 and KDM3A null cilia to CK-869 - KDM3Anull CK-869)Image 641833 (Figure 6G: Effect of ARP2/3 inhibition on IFT in RPE1 WT cells - RPE1 Wild Type Cell)Christopher PlayfootIan R. AdamsPatricia L. YeyatiNick GilbertRachel SchillerJimi WillsGirish MaliAlex von KriegsheimAndrew FinchPleasantine MillIan J. JacksonAkane KawamuraVeronica van HeyningenIoannis KasioulisJuro SakaiChristopher J. SchofieldfalseKDM3A coordinates actin dynamics with intraflagellar transport to regulate cilia stabilityCilia assembly and disassembly are coupled to actin dynamics, ensuring a coherent cellular response during environmental change. How these processes are integrated remains undefined. The histone lysine demethylase KDM3A plays important roles in organismal homeostasis. Loss-of-function mouse models of <jats:italic>Kdm3a</jats:italic> phenocopy features associated with human ciliopathies, whereas human somatic mutations correlate with poor cancer prognosis. We demonstrate that absence of KDM3A facilitates ciliogenesis, but these resulting cilia have an abnormally wide range of axonemal lengths, delaying disassembly and accumulating intraflagellar transport (IFT) proteins. KDM3A plays a dual role by regulating actin gene expression and binding to the actin cytoskeleton, creating a responsive “actin gate” that involves ARP2/3 activity and IFT. Promoting actin filament formation rescues <jats:italic>KDM3A</jats:italic> mutant ciliary defects. Conversely, the simultaneous depolymerization of actin networks and IFT overexpression mimics the abnormal ciliary traits of <jats:italic>KDM3A</jats:italic> mutants. KDM3A is thus a negative regulator of ciliogenesis required for the controlled recruitment of IFT proteins into cilia through the modulation of actin dynamics.2017-02-28T11:29:13Z2018-11-29T11:29:13Z2018-11-29T11:29:13ZS-JCBD-20160703210.1083/jcb.201607032