FIGURE 6:

FHDC1-induced cilia elongation is F-actin dependent. mCherry, FHDC1, or FHDC1.I180A (red) were expressed in NIH 3T3 cells by transient transfection, and the transfected cells were serum-starved to induce cilia assembly. At 40 h after transfection the cells were treated with 2-μm latrunculin B (Lat B) for 2 h and the effects on cilia length and number were assessed by immunofluorescence. F-actin (green) was detected with Alexa488-phalloidin and acetylated microtubules (white) were detected with anti-acetyl-α-tubulin. For all samples, LatB treatment did not affect the number of ciliated cells. (A, B) LatB treatment did not affect the length of cilia in mCherry-expressing cells. LatB treatment significantly reduced the length of cilia in FHDC1-expressing cells. LatB had minimal effects on cilia length in FHDC1.I180A-expressing cells. (C, D) Quantification of data shown in A and B. N = 3, >100 cells counted per sample. Error bars = SEM. Red bar indicates average cilia length.