Fig. 3: Functional analysis of the CEP128R222Q variant in hRPE1 cells.
From: CEP128 is involved in spermatogenesis in humans and mice
a Western blot analysis showed an increased abundance of CEP128 protein with the CEP128R222Q variant. NC (negative control), cells transfected with negative control pENTER-Flag plasmid. Three independent experiments were performed. (Two-sided Student’s t test; error bars, mean ± SEM). The p values are indicated in the graphs. b Overexpression of CEP128R222Q compromised ciliary growth and disrupted cellular nuclear morphology. Three independent experiments were performed. Dotted box denoted the basal body and cilia. n the number of cells analyzed. NC (negative control), cells transfected with negative control pENTER-Flag plasmid. (Two-sided Student’s t test; error bars, mean ± SEM; green, CEP128; red, Ac-Tubulin; blue, DAPI; scale bars, 10 µm). The p values are indicated in the graphs. c Decreased levels of NIN, centriolin, and CEP170 were detected in the cells transfected with WT-CEP128 or CEP128R222Q when compared to the NC by western blot analysis. NC (negative control), cells transfected with negative control pENTER-Flag plasmid. Three independent experiments were performed. (Two-sided Student’s t test; error bars, mean ± SEM). The p values are indicated in the graphs. WT-CEP128 and CEP128R222Q downregulated the mRNA levels for NIN, centriolin, and CEP170 (d) by suppressing the expression of their transcription factors (e). NC (negative control), cells transfected with negative control pENTER-Flag plasmid. Three independent experiments were performed. (Two-sided Student’s t test; error bars, mean ± SEM). The p values are indicated in the graphs. Source data are provided as a Source Data file.
