Fig. 1.

Various pro-inflammatory cytokines elongate primary cilia permanently. (a) TNFα was applied to MEC in different concentrations for 24 h (n ≥ 100 per concentration). (b) Time course of cilia elongation was quantified in MEC after addition of 10 ng/mL TNFα (n ≥ 43 cilia per time point). (c) Cilia length was determined after an initial trigger of 8h TNFα (10 ng/mL) followed by additional stimulation or an alternative wash step after 16 h, 24 h, or 48h. All conditions were fixed and quantified after 56 h (n ≥ 34 cilia per condition). (d) Maximum intensity projections (MIP) of representative (flat) cilia (arrow heads) upon 24 h treatment with TNFα (10 ng/mL), IL1β (10 ng/mL), IFNγ (100 U/mL), or the positive control FK (100 μM) (scale bar = 5 μm). Schematic depicts differences between MIP used to determine xy-length and three-dimensional (3D) length calculation applied for cilia length analysis and statistics. Quantification of average 3D cilia length is shown to the right (n ≥ 53 per condition).