Fig 2. Tissue-specific expression of MRO isoforms.

RT-PCR using MRO primer combinations demonstrated the presence of multiple splice variants, confirming the inferred sequences, and revealing a novel isoform (MROa2). (A) Schematic representation of the MRO RT-PCR primer annealing location. (B-i) PCR using primers set p1F-p9R, detected two products in testes and GCs (MROb2 and MROb4). A third isoform, MROb3, was cloned in GCs only. (B-ii) PCR using primers p3/5F-p9R detected MROa variant in the ovary and MROc and MROd in liver, kidney and brain. In the testes and GCs, this primer set detected MROb3 and b4. (B-iii) PCR using primer set p4F-p9R, detected MROa (all tissues) and MROa2 (brain) products. (B-iv) Human actin-B (hACTB) was used as loading control. NTC–RNA template control. Cycling conditions were as follows: 3 min at 95°C following by 40 cycles of 95°C/30 sec, annealing at 60°C/ 30 sec and extension at 68°C/ 1–2 min (contingent on the size of the expected amplification product).