Fig. 2: Gas5 is highly enriched in the synapse following fear extinction training.
From: Fear extinction is regulated by the activity of long noncoding RNAs at the synapse
a Illustration of alternative splicing (AS) events. b Splicing patterns of AS events in synapse- and nucleus- enriched lncRNAs. Skipped exons (synapse = 32%; nucleus = 17%; two-proportions Z-test, p = 2.41e−45), intron retention (synapse = 8%; nucleus = 21%; two-proportions Z-test, p = 2.13e−52). c Heatmap showing Gas5 isoforms expression generated using IsoVis. Red and green indicating high and low expression, respectively. Arrow indicates Gas5 variant ENSMUST00000162558.7. d RT-qPCR of Gas5 variant expression in the nuclear and synaptic fractions of the ILPFC. The amplified Gas5 exonic (E) and intronic (I) regions are indicated. Gas5 transcripts with and without introns are also indicated. Statistical significance was determined using two-tailed unpaired Student’s t test on independent biological replicates (E11-I11-E12, nucleus, n = 12, synapse, n = 10, t(11.02) = 16.91, p < 0.0001; I9-E10, nucleus, n = 6, synapse, n = 4, t(5.031) = 7.704, p = 0.0006; I8-E11, nucleus, n = 6, synapse, n = 4, t(5) = 8.452, p = 0.0004; E2-I2, nucleus, n = 6, synapse, n = 4, t(5.004) = 8.159, p = 0.0004; I5, nucleus, n = 6, synapse, n = 4, t(5.022) = 7.948, p = 0.0005; E1-E2-E3, nucleus, n = 12, synapse, n = 9, t(8.01) = 4.774, p = 0.0014; E8-E12, nucleus, n = 12, synapse, n = 9, t(8,083) = 3.834, p = 0.0049). ***p < 0.005, ****p < 0.0001. Error bars represent S.E.M. e RT-qPCR of Gas5 variant ENSMUST00000162558.7 in the nucleus and synapse fractions of ILPFC samples. Statistical significance was determined using two-tailed unpaired Student’s t test on independent biological replicates (Gas5, nucleus, n = 6, synapse, n = 9, t(8.05) = 3.453, p = 0.0086; Meg3, nucleus, n = 8, synapse, n = 6, t(7) = 9.763, p < 0.0001). **p < 0.01, ****p < 0.0001. Error bars represent S.E.M. f Representative image showing the localized expression of the Gas5 variant in primary cortical neurons (n = 8 fields of views). Arrowheads show synaptic localization. Scale bar, 20 µm. Red, Gas5; blue, DAPI; green, MAP2 protein. The boxed region is enlarged in the inserts. Scale bar, 5 µm.
