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. 2017 Jun 22;45(15):9093–9107. doi: 10.1093/nar/gkx536

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© The Author(s) 2017. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com

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Figure 7. — Functional overlap of alg-5, alg-1 and alg-2. (A) Western blot assay of HA::ALG-2 derived from a chimeric construct containing alg-1 5′ and 3′ regulatory sequence and alg-2 coding sequence (alg-1::HA::alg-2). Actin is shown as a loading control. (B) Proportion of burst or dead animals. Wild type (n = 152), alg-1(gk214) (n = 114), alg-1::HA::alg-1; alg-1(gk214) (n = 116), and alg-1::HA::alg-2; alg-1(gk214) (n = 92) animals were grown at 20°C. (C) Overlap of miRNAs enriched >1-fold (left) and >2-fold (right) in co-IP of HA::ALG-2 derived from alg-1::HA::alg-2; alg-1(gk214) with miRNAs uniquely enriched in co-IPs from HA::ALG-1 (alg-1::HA::alg-1; alg-1(gk214)) or HA::ALG-2 (alg-2::HA::alg-2; alg-2(ok304)) co-IPs >1-fold (left) and >2-fold (right). (D) Western blot assay of HA::ALG-5 derived from a construct containing the authentic alg-5 regulatory elements in the alg-5(tm1163) mutant background (alg-5(tm1163); alg-5::HA::alg-5) and a chimeric construct containing alg-1 5′ and 3′ regulatory sequences and alg-5 coding sequence in the alg-1(gk214) mutant background (alg-1::HA::alg-5; alg-1(gk214)). HA::ALG-1 from alg-1::HA::alg-1 in the alg-1(gk214) mutant background is also shown. Actin is shown as a loading control. glp-4 RNAi was done to reduce germ cell proliferation during development. L4440 vector RNAi was done as a control. (E) A developmental time course of HA::ALG-5 from alg-1::HA::alg-5; alg-1(gk214) (upper panel) and alg-5(tm1163); alg-5::HA::alg-5 (lower panel) and HA::ALG-1 from alg-1::HA::alg-1; alg-1(gk214) (middle panel). Actin is shown as a loading control. Numbers below blot images are signal intensities of HA normalized to actin (embryo samples arbitrarily set to 1.0). (F) Proportions of animals containing protruding or burst vulvas. Error bars represent standard deviations from the means from two independent experiments. Wild type (n = 104–124), alg-1(gk214) (n = 102–109), alg-1::HA::alg-1; alg-1(gk214) (n = 109–114) and alg-1::HA::alg-5; alg-1(gk214) (n = 102–116) animals were grown at 25°C See also Supplementary Figure S6.