MOESM3 of Absence of M-Ras modulates social behavior in mice

Additional file 3: Figure S3. (A, B) M-Ras protein expression in mouse tissues. (A) M-Ras expression was very high in brain (cortex), low in bladder, kidney, lung, spleen, and thymus, and undetectable in other tissues, including heart and skeletal muscle. (B) M-Ras protein expression in mouse brain regions. This Western blot shows highest expression of M-Ras in the cerebral cortex (COR) and lower expression in hypothalamus (HYP), hippocampus (HIP), and in the olfactory bulbs (OB). Note the absence of M-Ras in Mras −/− (KO) samples. (C) NS size frequency distributions at varying concentrations of EGF. Neural progenitor cells from SVZ tissue were cultured in the presence of different concentrations of EGF, as indicated. NS diameters (in µm) were measured at DIV7 (n = 6 for 0.2 ng/mL EGF, n = 5 for 2 ng/mL EGF, n = 3 for 20 ng/mL EGF; error bars represent SD). (D) Secondary NS growth. NSC from DIV7 NS grown in 2 ng/mL EGF were dissociated and replated at varying concentrations of EGF. To estimate cell proliferation, metabolic activity was measured six days after seeding by WST-1 assay (n = 3 for both WT and Mras −/−; error bars represent SD; significant effect of EGF stimulation on growth (F4,20 = 90.42, p < 0.0001); no significant effect of genotype. (E) NS size frequency distribution with and without PRL. Neural progenitor cells were cultured in the presence of 2 ng/mL EGF and with or without 2 ng/mL sheep PRL (sPRL; left) or 2 ng/mL recombinant murine PRL (rmPRL; right). NS diameters were measured at DIV7. Virtually identical results were obtained with sPRL and rmPRL. (F) PRLR gene expression. Levels of the three transcript variants of PRLR were quantitated by real-time PCR in SVZ tissue and in NSC from DIV7 NS. No PRLR was detected in cultured primary NS (n = 3 for all samples; data shown with SD).