Figure 2

Knockdown of neuroligin-2 abolishes GABA functional switch in cortical neurons. A, Representative images showing 8 DIV NLmiR-transfected neurons loaded with Fura-2. Somata of both nonTF and transfected neurons were selected (Fura-2 panel, white circles) to measure 340/380 ratio signal. Scale bar, 40 μm. B, Sample traces showing somatic Ca²⁺ responses after stimulation with 100 μM GABA and 90 mM KCl in nonTF neurons (gray, 8 DIV) and NLmiR-transfected neurons (black). C, Averaged sample traces showing that BIC (20 μM) blocked GABA-evoked Ca²⁺ responses in both nonTF (gray, n = 6) and NLmiR-transfected neurons (black, n = 17). D, The time courses of GABA functional switch in nonTF neurons and neurons transfected with mCherry, NLmiR, NLmiR + NL1*, NLmiR + NL2*, or NL2shRNA (3–4 independent cultures; n = 2291, 272, 433, 350, 322, 152 neurons, respectively). E, Bar graphs showing the amplitude of GABA-evoked Ca²⁺ increases in neurons transfected with mCherry, NLmiR, NLmiR + NL1*, NLmiR + NL2*, or NL2shRNA at 4 and 12 DIV. There was no difference between all groups at 4 DIV (p > 0.6). ***p < 0.001, one-way ANOVA. F, Percentage of mature neurons showing GABA-evoked Ca²⁺ increase after NLmiR or NL2shRNA transfection. Arrows in D and F indicate the time of transfection (TF).