Figure 5.

Lentiviral-based system for site-specific knockdown of CRFR2. A, Four different shRNA target sequences from the open reading frame of the mouse CRFR2 gene were designed. B, Western blot analysis followed by densitometry measurements show the ability of viruses #2 and #3 to reduce levels of CRFR2 expression in CRFR2 HEK293T transfected cells. C, The ability of shCRFR2 to decrease the functionality of the receptor was confirmed by assessing cAMP activation, after 4 h treatment with different concentrations of Ucn2. D, Site-specific delivery of shCRFR2#3 into the lateral septum. Representative in situ hybridization images of coronal brain sections of control (left) or unilaterally injected (right) mouse brain.