Figure 1. rRNA targets equally well to SN branches contacting target and non-target MNs.
A bifurcated SN was cultured with an L7 target MN and an L11 non-target MN (A) for 3 days. Alexa647 (cyan) was microinjected into the SN before imaging to visualize SN processes. Cultures were processed for Fluorescent In Situ Hybridization (FISH) for 18S rRNA (B). Areas in proximal SN neurites outlined with white squares in (A) were imaged at high magnification in (B); left panels show merged images of DIC and Alexa fluorescence, and right panels show FISH signals. The Alexa647 served as a volume control to analyze FISH RNA intensity. Group data show that 18S rRNA (C) is evenly distributed in neurites contacting L7 target and L11 non-target MNs. Parallel experiments showing FISH for 28S rRNA are shown in Figure 1—figure supplement 1. A bifurcated SN was cultured with an L7 target MN and an L11 non-target MN (D) for 1 day and a plasmid expressing ApS6 tagged with dendra2 was microinjected together with Alexa647 (cyan) into the SN. Live imaging (D) was performed on day 3; shown in (E) are high magnification images of DIC and Alexa fluorescence on the left and ApS6-dendra2 on the right. The Alexa647 served as a volume control to analyze FISH RNA intensity. Group data show that ApS6 (F) is evenly distributed in neurites contacting L7 target and L11 non-target MNs. Error bars represent SEM. None of the differences were significant as determined by a Student's paired t-test. Scale bar in (A and D) =100 μm; in (B and E) =20 μm.
Figure 1—figure supplement 1. 28S rRNA targets equally well to SN branches contacting target and non-target MNs.
