Fig. 2
Western Blot analysis for calretinin and RT-PCR for the detection of Calb2 mRNA. a) Western blot signals (upper panel) and the corresponding Ponceau Red-stained membrane (lower panel). As positive control, 10 ng of purified human recombinant CR was used. Sizes (molecular masses) of the marker proteins in the right lane are (from top to bottom): 130, 100, 75 (red), 63, 48, 35, 28 (green), and 17 kDa. The specific signal for CR runs just slightly above the green marker protein at approximately 30 kDa. In MSTO-211H cell extracts, a strong signal at the same position as purified CR indicates the presence of CR in these cells. All mouse samples from the MM cell lines RN29, RN5 and AK7 (all from C57Bl/6 J mice), AB12 (Balb/c) are negative, as well as the negative control prMC from a CR−/− mouse (prMC−/−). Mouse cerebellum extract (most right) shows a strong signal for CR. b) Positive signals for Calb2 mRNA were detected in RN29, RN5, AB12, AK7 and iMeso-WT cells; no signal was present in samples from freshly isolated C57 mouse prMC and in the negative control (H2O). RPS13 was used as housekeeping gene