Fig 1: In vivo treatment efficacy of AAV8-HLP-mGHR in Laron mice.a Timeline summarized the experiment performed. b Representative photos for uninjected control mice (GHR+/+, GHR+/− and GHR−/−) and Laron mice (GHR−/−) injected with 4 × 1010 vg/mouse of AAV8-HLP-Luc and AAV8-HLP-mGHR in male (i) and female (ii). Original unmodified image was shown in Supplementary Fig. S1. Weekly measurement of (c) body weight and (d) body length post AAV injection and the corresponding (e) normalized body weight and (f) normalized body length. Body weight and length were normalized to 100% at the time of AAV injection to correct the initial body weights and length variation of mice. AAV8-HLP-mGHR and AAV8-HLP-Luc treated groups were monitored for 25–26 weeks post AAV injection while uninjected control groups were monitored for 16 weeks only. Data were presented as mean ± s.e.m. n = 5–6. Statistical significances were determined by two-way ANOVA where **** indicates p < 0.0001 and considered extremely significant.
Fig 2: Circulating GH, IGF1, ALS and IGFBP3 of AAV8-HLP-mGHR treated Laron mice.Serum level of mouse (a) GH, (b) IGF1, (c) ALS, (d) IGFBP3 and (e) IGF1:IGFBP3 molar ratio at end point in (i) male and (ii) female of uninjected GHR+/+, GHR+/−, GHR−/− and AAV-injected Laron dwarf mice with AAV8-HLP-Luc and AAV8-HLP-mGHR. Data were presented as mean ± s.e.m. n = 3–6. Statistical significances were determined by Student’s unpaired t-test where ns indicated not significant, * indicates p < 0.05 and considered statistically significant, ** indicates p < 0.01 and considered very significant, *** indicates p < 0.001 and considered extremely significant and **** indicates p < 0.0001 and considered extremely significant.
Fig 3: Effect of AAV8-HLP-mGHR treatment on femurs length and organs sizes of Laron dwarf mice.a Femur length in (i) male and (ii) female of uninjected GHR+/+, GHR+/−, GHR−/− and AAV-injected Laron dwarf mice with AAV8-HLP-Luc and AAV8-HLP-mGHR. Data were presented as mean ± s.e.m. n = 4–6. Statistical significances were determined by Student’s unpaired t-test where ns indicated not significant and **** indicates p < 0.0001 and considered extremely significant. b Photo of organs and femurs of AAV8-HLP-Luc and AAV8-HLP-mGHR injected Laron dwarf mice harvested at endpoint. Original unmodified image was shown in Supplementary Fig. S1.
Fig 4: Confirmation of viral gene delivery to liver and its effect on mouse transcript of GHR, IGF1, ALS and IGFBP3.a Quantification of viral transgene copies number in mouse liver. The copies numbers were normalized with mouse GAPDH housekeeping gene. Messenger RNA expression of mouse (b) GHR, (c) IGF1, (d) ALS and (e) IGFBP3 at end point in (i) male and (ii) female of uninjected GHR+/+, GHR+/−, GHR−/− and AAV-injected Laron dwarf mice with AAV8-HLP-Luc and AAV8-HLP-mGHR. The mRNA expression levels were normalized with 18S rRNA housekeeping gene. Data were presented as mean ± s.e.m. n = 3–6. Statistical significances were determined by Student’s unpaired t-test where ns indicated not significant, * indicates p < 0.05 and considered statistically significant, ** indicates p < 0.01 and considered very significant, *** indicates p < 0.001 and considered extremely significant and **** indicates p < 0.0001 and considered extremely significant.
Supplier Page from Sino Biological, Inc. for Mouse Growth Hormone Receptor / GHR / GHBP transcript variant 1 Gene ORF cDNA clone in cloning vector