Analysis of the efficacy of NvStbm knockdown using antisense NvStbm morpholinos. (A) Western blot analysis was used to determine the efficacy of the NvStbm-MO. Individual protein band pixel intensities were measured using the Odyssey software (LI-COR, Lincoln, NE, USA). These intensities were used to calculate the ratios of NvStbm/ß-tubulin and normalized to determine the fold difference of each NvStbm band. This analysis showed that there is approximately a five-fold decrease of the endogenous Stbm protein in NvStbm-MO-injected (650 μM) embryos compared to embryos injected with the Control-MO (650 μM). (B, C) Scanning confocal microscopical analysis of Control- and NvStbm-MO injected embryos immunostained with affinity-purified NvStbm antibodies. (B) NvStbm antibody stained Control- MO-injected (650 μM) embryos showed strong NvStbm expression at the apical end of invaginating cells at the blastopore while (C) a majority of NvStbm-MO (650 μM) injected embryos did not show NvStbm expression. Also, NvStbm-MO injected embryos did not undergo initial archenteron invagination. The numbers in (B) represent the number of cases with NvStbm staining, and the numbers in (C) represent the number of cases without NvStbm staining.