Figure EV5. Fam134b‐HA mRNA expression rescued Fam134b ^mo skeletal phenotype.

• A
  qRT–PCR analysis of Fam134b gene in medaka fish with indicated genotypes. Values were normalized to Hprt gene and expressed as fold change relative to scramble fish. Mean ± standard error of the mean (SEM) of N = 3 biological replicates.
• B
  Reverse transcription–PCR analysis of Fam134b gene in Fam134b ^wt and Fam134b ^mo medaka fish. Actin gene was used as control gene. M = marker; B = blank.
• C
  Western blot analysis of HA‐tag from a pool of medaka fish embryos injected with scramble or injected with mRNA produced from human HA‐FAM134B pcdna3.1(+). β‐actin was used as a loading control.
• D
  Bar graphs show quantification of total length and head size of medaka fish model of Fam134b ^mo and mRNA‐injected Fam134b ^mo expressed as % relative to the scramble. Mean ± standard error of the mean (SEM) of at least n = 8 fish per genotype. Student's unpaired t‐test *P < 0.05; NS, not significant.
• E, F
  Bar graphs show quantification of Alcian Blue (cartilage) (e) and Alizarin Red (bone) (f) staining of Fam134b ^mo and mRNA‐injected Fam134b ^mo. Ethmoid plate (EP), palatoquadrate (PQ), ceratohyal (CH), paired prootics (PO), ceretobranchials 1–5 (CB1 to CB5) cartilage length (E), and bone mineralization (F) were evaluated. Values were expressed as % relative to the scramble (100% red dotted line). Mean ± standard error of the mean (SEM) of at least n = 6 fish/genotype. Student's unpaired t‐test *P < 0.05; **P < 0.005; ***P < 0.0005 for comparison between Fam134b ^mo and mRNA‐injected Fam134b ^mo medaka.

Source data are available online for this figure.