Journal: Scientific Reports

Article Title: Reduced IGF-1 signaling fails to limit Alzheimer’s disease progression in a novel rat model of IGF-1R haploinsufficiency

doi: 10.1038/s41598-025-31601-1

Figure Lengend Snippet: Characterization, and validation of Igf1r heterozygous rats. ( A , B ) We first confirmed that the 1 bp-C insert resulted in a functional Igf1r knockout on one allele by performing western blotting of IGF-1R in several tissues. These studies revealed that IGF-1R expression was reduced by > 50% in lung, heart, hypothalamus and kidney of male rats (n = 4 per group). ( D ) As a positive control, InsR was also measured in the same samples and confirmed to not be altered in animals. ( C – F ) To further confirm that igf1r heterozygosity results in attenuated signaling, we treated dermal fibroblasts isolated from young control and Het rats and administered either Vehicle or 25 nM IGF-1 for 10 min. As can be seen, Igf1r Het fibroblasts demonstrate an attenuation in activation of pAkt, but not pErk and pS6 under IGF-1 stimulated conditions ( C – F ; p < 0.05). Statistics were performed either via independent samples t-test or two-way ANOVA, followed by posthoc tests with Tukey adjustment. Bar graphs represent mean ± S.E.M. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Article Snippet: Membranes were blocked in 5% milk or BSA in TBST and then incubated with appropriate primary antibodies from Cell Signaling Technologies (Danvers, MA) against IGF-1R (Cell Signaling; #9750), InsR (Cell Signaling; #3025), p-GSK3β (Cell Signaling; #5558), total GSK3β (Cell Signaling; #12456), p-Tau(Ser202/Thr205) (Thermo Fisher AT8 #MN1020), total Tau (Cell Signaling; #46687), amyloid-beta (Cell Signaling; #8243), pS6 (catalog #5364), total S6 (Cell Signaling; #2217), p-Erk (Cell Signaling; #9101), total Erk (Cell Signaling; #4695), p-MAP2 (Ser136) (Cell Signaling #4541), Total MAP2 (Cell Signaling; #8707) overnight at 4 °C overnight with gentle agitation.

Techniques: Biomarker Discovery, Functional Assay, Knock-Out, Western Blot, Expressing, Positive Control, Isolation, Control, Activation Assay