Fig. 1
From: Progranulin promotes peripheral nerve regeneration and reinnervation: role of notch signaling
Characterization of tamoxifen inducible neuronal progranulin overexpressing mice. SLICK-Grn-OE mice were generated by mating homozygous STOP-Grnflfl mice (progranulin cDNA with leading STOP sequence into Rosa26 locus) with SLICK-H Cre mice, resulting in overexpression of progranulin in the majority of central and peripheral neurons after tamoxifen treatment. a Quantitative RT-PCR of progranulin mRNA in dorsal root ganglia (DRGs), spinal cord, brain regions and motor cortex, the latter after sciatic nerve injury in SLICK-Grn mice (without tamoxifen, noTAM, = control) and SLICK-Grn-OE mice (with tamoxifen, TAM). Progranulin knockout (Grn−/−) mice show the primer specificity. The contralateral motor cortex is the cortical representation of the injured nerve. Each symbol is a mouse, the line is the mean and whiskers show the standard deviation (SD). b Enzyme immunoassay of progranulin in spinal cord, brain and primary culture supernatants in SLICK-Grn (without tamoxifen) and SLICK-Grn-OE (with tamoxifen) mice. Grn−/− mice show the antibody specificity. The figure shows scatter plots of individual mice with mean ± SD. Tamoxifen treatment significantly increased Grn mRNA and protein expression throughout regions (rm-ANOVAs for genotype P < 0.05) without significant differences between the ipsilateral and contralateral sides. c In situ hybridization of progranulin mRNA in the spinal cord, hippocampus and motor cortex of Grn−/− mice, STOP-Grnflfl mice, SLICK-Grn (without tamoxifen) and SLICK-Grn-OE (with tamoxifen) mice. d Immunofluorescence analysis of EYFP and progranulin in SLICK-Grn mice (without tamoxifen) and SLICK-Grn-OE mice (with tamoxifen) in the DRGs. The double-headed Thy1 promotor drives constitutive EYFP expression and inducible CreERT-dependent expression of Cre recombinase that cuts out the STOP codon in front of progranulin cDNA. Hence, progranulin expression increases after tamoxifen, EYFP remains constant. In situ and immunofluorescence images are representative results of 3–4 mice per group. Scale bars: 50 μm. e Immunofluorescent analysis of progranulin in injured (ipsilateral) and uninjured (contralateral) sciatic nerves in SLICK-Grn mice (without tamoxifen) and SLICK-Grn-OE mice (with tamoxifen). Full time courses are shown in Additional file 1: Figure S2. Scale bars: 50 μm