Skip to main content
. 2018 Sep 26;16(9):e2004874. doi: 10.1371/journal.pbio.2004874

Fig 3. mDia1/3-dependent rapid dynamics of cortical F-actin meshwork in primary cultured Sertoli cells.

Fig 3

(A) Time-lapse images of WT primary cultured Sertoli cell expressing LifeAct-EGFP. Red arrows indicate the tip of the elongating F-actin in the cortical meshwork. Time is shown in seconds. The colors in the right panel indicate the elongated actin filament at each time point in the left panel. Scale bar, 2 μm. (B) Quantification of the elongation speed of LifeAct-EGFP in WT and mDia1/3 DKO Sertoli cells. Results from two independent experiments. Data are represented as mean ± SEM (n = 49 and 20 filaments from four WT and four mDia1/3 DKO Sertoli cells, respectively). **P < 0.01 (P = 0.0013, Student t test). (C) Quantification of the straightness of the elongated F-actin in WT and mDia1/3 DKO Sertoli cells. Results from two independent experiments. Data are represented as mean ± SEM (n = 49 and 20 filaments from four WT and four mDia1/3 DKO Sertoli cells, respectively). (D) Quantification of the elongation frequency of cortical F-actin meshwork. Results from two independent experiments. Data are represented as mean ± SEM (n = 9 regions from three WT and three mDia1/3 DKO Sertoli cells, respectively). ***P < 0.001 (P = 0.0002, Student t test). (E) Time-lapse images of WT primary Sertoli cell expressing EGFP-mDia3. Red boxes indicate a single molecule of EGFP-mDia3. Time is shown in seconds. Scale bar, 2 μm. (F) Kymograph of EGFP-mDia3 single molecule in Fig 3E. (G) Quantification of the processive moving speed of EGFP-mDia3 single molecules (n = 59 molecules from three cells). The result is representative of two independent experiments. (H) Quantification of the straightness of EGFP-mDia3 single molecules’ processive movement (n = 59 molecules from three cells). The result is representative of two independent experiments. (I) Quantification of the travel distance of EGFP-mDia3 single molecules (n = 59 molecules from three cells). The result is representative of two independent experiments. DKO, double knockout; EGFP, enhanced green fluorescent protein; F-actin, filamentous actin; mDia1/3, mammalian diaphanous homolog1/3; n.s., not significant (P = 0.3194, Student t test); WT, wild-type.