C. elegans Developmental Proteome Analysis

C. elegans were synchronized at 20 °C by bleaching gravid adult animals and maintaining starved L1 larvae for at least 24 h before plating on OP50. For sample collection, animals were harvested after 2 h (L1), 12 h (L2), 32 h (L3), 50 h (L4), and 72 h (gravid adult) on OP50. Number of animals loaded per lane was normalized for actin—~1000 L1s, 800 L2s, 600 L3s, 400 L4s, and 200 gravid adults. Proteins were resolved on 4–12% Bis-Tris polyacrylamide gels (Thermo Fisher), transferred to nitrocellulose membranes (Thermo Fisher), and probed with rat anti-HA-peroxidase 1:1000 (Roche 12013819001), mouse anti-FLAG 1:1000 (Sigma, F1804), mouse anti-actin 1:10,000 (Abcam ab3280), or rabbit anti-CSR-1 1:2000 antibodies^{8 (link)}. Secondary HRP antibodies were purchased from Thermo Fisher. Unedited western blots provided in the Source Data File.

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Nguyen D.A, & Phillips C.M. (2021). Arginine methylation promotes siRNA-binding specificity for a spermatogenesis-specific isoform of the Argonaute protein CSR-1. Nature Communications, 12, 4212.

Publication 2021

4–12% Bis-Tris polyacrylamide gels nitrocellulose membranes rat anti-HA-peroxidase mouse anti-FLAG mouse anti-actin Secondary HRP antibodies

Actin Adults Animals Antibodies Bis tris C elegans Larvae Membranes Mouse Nitrocellulose Peroxidase Polyacrylamide gels Proteins Rabbit Sample collection Western blots

Corresponding Organization :

Other organizations : University of Southern California

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Variable analysis

independent variables

Time points (2 h (L1), 12 h (L2), 32 h (L3), 50 h (L4), and 72 h (gravid adult))

dependent variables

Protein expression levels of HA-tagged, FLAG-tagged, actin, and CSR-1

control variables

Temperature (20 °C)
Species (C. elegans)
Food source (OP50)
Number of animals loaded per lane (normalized for actin)

positive controls

Not explicitly mentioned

negative controls

Not explicitly mentioned

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