Fig. 1

Expression of Mb gene transcript variants and Mb protein in PAM-MSCs and MAT-MSCs. A Scheme showing the structure of the alternative transcripts of the Mb gene; variant 2 (NM_005368) and variant 13 (NM_203377). B Quantitative RT-PCR analysis of the transcript variants 2 (Mb-v2) and 13 (Mb-v13) in PAM-MSCs and MAT-MSCs normalized to their expression in rhabdomyosarcoma cells (RD); means ± SEM of 4–5 independent measurements (biological replicates) each carried out in 3 technical replicates; *, P < 0.05 and ***, P < 0.001 with respect to RD. C Relative amounts of Mb-v13 compared to that of Mb-v2 expressed as ratios of the relative 2^−ΔCt values; the values shown are from the measurements in (B); **, P < 0.01 with respect to RD. D Comparative immunoblot for Mb detection. 40 µg protein extract from total cell lysates of four and three different preparations of PAM-MSC and MAT-MSC, respectively, was assayed by SDS-PAGE and Western blotting as described in Materials and Methods; 40 µg protein extract from rhabdomyosarcoma-derived cell line (RD) and 0.2 µg pure horse heart myoglobin (Pur. Mb) were co-analyzed for comparative purpose. E Densitometric analysis of comparative WBs as that shown in (D); the values refer to the 35 KDa and 17 kDa band, corresponding to the dimeric and monomeric Mb, respectively, normalized to the GAPDH band and to the value observed for the RD cells; means ± SD of six different preparations of both PAM- and MAT-MSCs; NS, not statistically significant