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. 2016 May 5;6(6):798–805. doi: 10.1016/j.stemcr.2016.04.004

Figure 4.

Figure 4

Disruption of Cilia Reassembly Impairs Satellite Cell Self-Renewal

(A) Schematic representation of the experimental design used in (B–E).

(B) Representative images of EDL T72 myofiber cultures treated with DMSO (control), taxol (2 μM), nocodazole (4 μM), or forchlorfenuron (FCF, 10 μM) at T66, and analyzed by immunofluorescence for the presence of primary cilia (ARL13B; green) in SCs (CAVEOLIN-1, red). White arrowheads indicate the presence of cilia.

(C) Representative images of EDL myofibers cultured in the presence of DMSO (control) or nocodazole or FCF and analyzed for the incorporation of EdU. A graph shows the percentage of EdU-positive cells per myofiber in each culture condition. n = 18–23 fibers. ns, not significant.

(D) Representative images of EDL myofibers cultured in the presence of DMSO (control), nocodazole, taxol, or FCF and analyzed by immunofluorescence using antibodies against PAX7 (green), MYOGENIN (magenta), and M-CADHERIN (red). Nuclei were counterstained with DAPI (blue).

(E) Percentage of cells observed in control cultures and in cultures following treatment with taxol, nocodazole, and FCF. Note that self-renewing SCs are PAX7+/MYOGENIN− (dark gray). n = 50–85 fibers from three independent experiments analyzed per condition; mean ± SEM is shown. ∗∗∗p < 0.0001, ∗p < 0.05.