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. Author manuscript; available in PMC: 2021 Aug 1.
Published in final edited form as: Dev Biol. 2020 May 20;464(1):35–44. doi: 10.1016/j.ydbio.2020.05.005

Figure 2. Knocking down of Oc, Sox102F and Ets65A using CRISPR-Cas9 system.

Figure 2.

(A) Oc expression (cyan) in control medulla (genotype T1LexA>LexopRFP/ UASCas9; eyGal4/+) with T1 labeled in red. (B) Oc expression (cyan) in ocgRNA medulla (genotype T1LexA>LexopRFP /UASCas9; eyGal4/ocgRNA) with T1 labeled in red. (C) Quantification of percentage of T1 neurons stained positive for Oc in control and ocgRNA brains, n=4 for control, n=8 for ocgRNA. Less than 100% ( ~93±1.8 %) of wild types T1s are stained positive for Oc, possibly due to focal planes or weak antibody signals, etc. Oc is lost in ~59±13 % of T1 neurons when Oc is knocked down with ocgRNA (t-test: p=6.24×10−6 ). (D) Sox102F expression (cyan) in control medulla (genotype T1LexA>LexopRFP/ UASCas9; eyGal4/+) with T1 labeled in red. (E) Sox102F expression (cyan) in Sox102FgRNA medulla (genotype T1LexA>LexopRFP/ UASCas9; eyGal4/Sox102FgRNA) with T1 labeled in red. (F) Quantification of percentage of T1 neurons stained positive for Sox102F in control and Sox102FgRNA brains. n=4 for control, n=9 for Sox102F-gRNA. Sox102F is lost in ~52±9% of T1 neurons in Sox102FgRNA brains. (t-test: p=8.39×10−8 ). (G–G”) In clones (marked in green) where Cas9 and Ets65AgRNA are expressed (genotype yw hs FLP; act>y+>Gal4 UAS GFP / UASCas9; UASEts65AgRNA/+), in situ hybridization of Ets65A (purple) in 3rd instar larval medulla showed that Ets65A mRNA is partially knocked down in the clones. Note the clone margin perfectly correlates with that of higher Ets65A expression.