Fig. 4

xtCYTC transcripts, xtCYTC protein, and xtCOXIV enzymatic activity expressions under various environments. (a) Tissue distribution of xtCYTC transcripts in various tissues collected from habitat xtcrabs and aquarium xtcrabs (n = 6 crabs/group). aGi, anterior gill; pGi, posterior gill; St, stomach; DG, digestive gland; In, intestine; Mu, muscle; He, heart. (b) xtCYTC transcripts in pGi of habitat xtcrabs and aquarium xtcrabs (n = 36 crabs/group). (c) xtCYTC protein expression as measured by Western blot (with xtCYTC antiserum) in the pGi of habitat and aquarium xtcrabs. Selected blot images are depicted above the graphs. Actin was used as a reference protein. Quantification of signal intensity was performed with ImageJ software (n = 3 xtcrabs/group). (d) COXIV enzymatic activity in habitat xtcrabs and aquarium xtcrabs (n = 6 crabs/group). Different letters indicate one-way ANOVA in xtCYTC tissue distribution (p < 0.05). EF1A was used as an internal reference to normalize the gene expression levels. The highest value among groups is set at 100, and each value is normalized to the ratio (each value/highest value) x100. Asterisks indicate significant difference between groups by t-test (*: p < 0.05; **: p < 0.01; ***: p < 0.001). The bars represent mean ± SD (standar deviation).