Fig. 3

Mutations in four Chinese families and their pathogenic effects. a 3D modeling of mutant proteins using SWISS-MODEL to illustrate structural alterations. b Anti-FLAG western blot assay demonstrating truncation of mutant proteins p.T706DfsTer52 and p.A715VfsTer31 at approximately 140 kDa, whereas the mutant protein p.L46V exhibits a size similar to the wild-type at 245 kDa. c TCOF1 mRNA expression in patient (MT) and healthy family member as control (WT). d Subcellular distribution of treacle indicated by anti-FLAG immunofluorescence. Scale bar = 100 μm. Each experiment was repeated at least three times. e TCOF1 mRNA overexpression validated by qPCR in HEK293T cells. f The relative expression of 45S rRNA validated by qPCR in HEK293T cells. P values obtained from two-tailed Student’s t test. n = 3 biologically independent samples. Error bars represent standard errors. Significance levels are indicated as *, P < 0.05; ***, P < 0.001