mRNA expression profiles of genes related to tight junction components and intermediate filaments.

Testicular transcript levels for Cldn11 (panel A), Cldn3 (panel B), Vim (panel C), Cdh2 (panel D), Espn (panel E), ZO-1 (panel F), Cx43 (panel G), Ocln (panel H) and Jam3 (panel I) were measured by qPCR and corrected for exogenously added luciferase mRNA as described in the Methods section. qPCR data (bars) represent the mean ± SEM of measurements on 3 SCARKO and 3 control testes studied on day 8, 10, 12, 15, 20, 25 and 35 postnatal (p.n.). The mean transcript level of the control measured on day 10 was arbitrarily assigned a value of 100. Statistically significant differences between SCARKO and control animals are indicated by an asterisk. The original microarray data (lines) are shown for comparison. Microarray measurements were performed on a pool of mRNA derived from testes of 3 SCARKO and 3 control mice (day 8, 10, 12, 16, 20 p.n.) [19]. Values (relative signal intensities) were expressed relative to the value of the control on day 10 (arbitrarily assigned a value of 100). Note that microarray measurements reflect transcript levels in a given amount of RNA. Due to the increasing contribution of germ cells selectively in control mice and not in SCARKO mice (which develop a block in meiosis), SC transcripts become diluted in the total amount of RNA and microarrays may fail to demonstrate differences in the expression of SC genes after day 15. The correction of the qPCR data for exogenously added luciferase circumvents this problem by reflecting transcript levels per testis [19].