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. 2018 Nov 29;131(23):jcs222042. doi: 10.1242/jcs.222042

Fig. 1.

Fig. 1.

Tpm3.5 mRNA and protein are reduced in Tpm3/Δexon9d−/− mouse lenses. (A) Diagram of Tpm3 exons. Alternative splicing produces six known Tpm3 isoforms in mice. Not drawn to scale. (B) RT-PCR for Tpm3 isoforms in cDNA from Tpm3/Δexon9d+/+ and Tpm3/Δexon9d−/− lenses and brain reveal that Tpm3 exon 9d is not detected in the lens, and instead contain Tpm3 exon 9a whose levels are unexpectedly reduced in the 9d-knockout (Tpm3/Δexon9d−/−) lens. Brain samples confirm that exon 9d is deleted. A diagram of the exons in Tpm3.5 is shown underneath. (C) Semiquantitative RT-PCR from three separate lens Tpm3/Δexon9d+/+ and Tpm3/Δexon9d−/− cDNA samples reveals decreased Tpm3.5 transcript levels. G3DPH was used a housekeeping gene and loading control. (D) Western blots of Tpm3/Δexon9d+/+ and Tpm3/Δexon9d−/− lenses reveal significantly decreased levels of Tpm3.5. Tpm3.1 and Tpm3.2, containing exon 9d, is not detected in lens samples. GAPDH is shown here to demonstrate equal loading of the samples. Ponceau S staining of total proteins on blots was used as a loading control. Plots reflect mean±s.d. of n=3 independent samples per genotype. **P<0.01.