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Data S1. Supplementary methods.

Figure S1. Blocking the β1-ADR with betaxolol (1 mg/kg) impaired social recognition independently of nonsocial odor recognition and object recognition. (A) In the three-chamber social recognition test, systemic injection of betaxolol prior the learning phase of the test did not alter the preference of mice for an unfamiliar intruder in a cup over an empty cup (vehicle-injected mice n = 10, betaxolol-injected mice n = 10) (two-way ANOVA interaction: = 0.2801, F(1,36) = 1.203; object < 0.0001, F(1,36) = 33.05; treatment = 0.6501, F(1,36) = 0.2093. Post hoc paired one-tailed t-test corrected for multiple comparisons: vehicle = 0.0004; betaxolol = 0.0014. However, betaxolol impairs social recognition: betaxolol-injected mice did not show a preference for a new intruder over a familiar one while vehicle-injected mice show this preference (two-way ANOVA interaction: = 0.0906, F(1,36) = 3.024; object = 0.0293, F(1,36) = 5.155; treatment = 0.6749, F(1,36) = 0.1789. Post hoc paired one-tailed t-test correlated for multiple comparisons: vehicle = 0.0082, betaxolol = 0.7128). (B) Betaxolol did not impair nonsocial odor recognition; in a test of nonsocial odor habituation, dishabituation, and recognition, both vehicle-injected (n = 10) and betaxolol-injected (n = 10) mice habituated to nonsocial odors (water and vanilla) over the course of three successive exposures and dishabituated when presented to a new odor (two-way RM ANOVA: interaction: = 0.9623, F(6,108) = 0.2401; treatment = 0.5234, F(1,108) = 0.4236; object < 0.0001, F(6,108) = 21.42; Post hoc paired one-tailed t-test correlated for multiple comparisons: Water habituation: vehicle ***< 0.001; betaxolol τττP < 0.001; Water dishabituation: vehicle **= 0.0024; betaxolol τττP < 0.001; Vanilla habituation: vehicle ***< 0.001; betaxolol ττ= 0.006). All mice were also able to discriminate between a previously presented odor (vanilla) and a new odor (mint) as indicated by higher sniffing time when presented with the mint odor versus sniffing time when presented with the vanilla odor for a third time (vehicle **= 0.0048; betaxolol ττ= 0.0036) (C) The ability of mice to recognize a familiar object over a new one was not affected by betaxolol. Both vehicle-injected (n = 10) and betaxolol-injected (n = 10) mice preferred a new object over a familiar one in an object recognition test (two-way ANOVA interaction: = 0.7487, F(1,36) = 0.1042; object < 0.0001 F(1,36) = 28.12; treatment P = 0.2604, F(1,36) = 0.2604; Post hoc paired one-tailed t-test correlated for multiple comparisons: vehicle < 0.001; betaxolol < 0.001). Data are presented as mean ± SEM.

Figure S2. (A) c-Fos expression is induced in the MeA 90 min after social recognition. Quantification of c-Fos positive cells was performed using a nonbiased stereological methods in control mice (n = 4) and mice exposed to a social recognition task (n = 4) (= 0.0303 by t-test). (B) c-Fos expression was not induced in the basolateral amygdala (BLA) after social recognition. Quantification of c-Fos positive cells was performed using a nonbiased stereological methods in control mice (n = 4) and mice exposed to a social recognition task (n = 4) (= 0.4857 by Mann–Whitney test). (C) Injection of betaxolol prior to testing in a social recognition task did not affect the number of c-Fos positive cells in the thalamus (PV), brain region not involved in social memory and poor in β1-ADR (vehicle-injected mice n = 4; betaxolol-injected mice n = 4; = 0.8571 by Mann–Whitney test). Data are presented as mean ± SEM. Scale bars, 100 µm (magnified box) and 200 µm.

Figure S3. Western blot analysis of pCREB performed on whole tissue homogenates from medial amygdala of control mice and APP mice (n = 4 per group). pCREB level was significantly higher in APP mice (= 0.0286 by Mann–Whitney test). The relative optical density is normalized to CREB. Data are presented as mean ± SEM.

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