Figure EV3. Projections from the primary motor cortex to CM and PCN.

• A, B
  Representative coronal images from the Allen Mouse Brain Connectivity Atlas (https://connectivity.brain‐map.org/projection/experiment/100141780?imageId=102152345), following anterograde labeling with AAV‐GFP in the primary motor cortex showing site of injection (A) and axonal distribution pattern in CM and PCN (B).
• C
  Representative coronal sections of the mPFC immunostained for c‐Fos with the anatomical borders of the ACC indicated by dashed lines, in virgin male mice on day 1 and 3 of behavioral testing. Scale bar 100 μm, 20× magnification. Z‐score for the difference in the fraction of c‐Fos‐expressing cells, between day 1 and 3 of behavioral testing.
• D
  Fraction of c‐Fos⁺ cells in the CL central lateral nucleus in virgin male mice exposed to pups, on day 1 and 3 of behavioral testing (Day 1: n = 4, Day3: n = 3). Data were analyzed using Student's t‐test and are expressed as mean ± s.e.m.
• E, F
  A representative image of the PFC of a virgin female following initial pup exposure showing mRNA expression patterns of slc17a7 (VGluT1), gad1 (GAD1), and c‐fos (c‐Fos) together with high‐magnification images of the ACC, following the 1^st and 3^rd days of pup‐exposure. The scale bars 500 μm, 10× magnification (A), and 100 μm, 20× magnification (B).
• G
  A comparison of the fraction of excitatory and inhibitory c‐Fos⁺ cells, of the total VGluT1 and GAD1 cell population in the ACC, following the 1^st and 3^rd pup exposure days; data were analyzed using Fisher's exact test; **P < 0.01.

Source data are available online for this figure.