Fig. 4.

Generation of brain-specific Igfbp2 knock-out (Igfbp2 cKO) mouse. (a) Construction of Igfbp2-floxed mouse. Red arrows show the genotyping primers, crRNA1 check primer F, and crRNA2 check primer R in Supplemental Table 2, to detect the Igfbp2 KO allele. (b) Sequence analysis to confirm the loxP knock-in. (c) PCR results to detect the Igfbp2 KO allele by Cre/loxP recombination. F1 mouse is Igfbp2^+/flox::CaMK2a-Cre, and the cont is the Igfbp2^+/flox mouse as a control. Bottom schema shows the construction of Igfbp2 KO allele primers. (d) Expression level of the Igfbp2 mRNA in the hypothalamus and liver of the male Igfbp2 cKO (n = 6) and the control mice (n = 3) exposed to 38 °C at their early stage of the embryo. Error bar shows SEM. * p-value < 0.05 (Student’s t-test).