(A) qPCR comparing the expression levels of Tmem176b during differentiation of Cry1 and Cry2 KD cells. Two shRNA clones were used for each gene. The expression level of Tmem176b in day 0 control cells was defined as 1.0.
(B) Temporal profiles of Tmem176b expression levels in WT, Cry1−/−, and Cry2−/− TA muscles.
(C) Temporal profiles of Tmem176b expression levels in C2C12 cells synchronized with forskolin between −1 and 0 hr and induced to differentiate at 0 hr. shRNA clone 1 was used for Cry1 and Cry2 KD.
(D) MHC staining of Tmem176b KD cells on differentiation day 5. Scale bar, 100 μm.
(E–G) Differentiation index (E), fusion index (F), and < 3 nuclei index (G) of Tmem176b KD cells on day 5.
(H) Frequency of nuclei in MHC(+) cells containing 1, 2, 3–5, 6–10, and more than 10 nuclei within one cell during differentiation. A total of 1,000 nuclei were counted at each time point.
Data are presented as mean + or ± SD.