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. 2013 Dec 19;289(6):3444–3456. doi: 10.1074/jbc.M113.507541

FIGURE 6.

FIGURE 6.

Effects of mutations in wdr-20 and wdr-48 on GLR-1::GFP accumulation and glr-1-dependent behavior. A, schematic of the structure of the coding and intronic sequences of wdr-20 (top panel) and wdr-48 (bottom panel). Locations of the mutations in gk547140, gk173034 and tm4575 are shown. Single nucleotide changes are indicated by arrows, and the deletion in tm4575 is denoted by a bracket. Schematics were generated using the Exon-Intron Graphic Maker (Wormweb). B, transgenic animals expressing the wdr-20 or wdr-48 transcriptional reporters Pwdr-20::GFP (left panel) or Pwdr-48::GFP (right panel), respectively, were imaged. The dotted white line outlines the worm body. Head neurons are marked by a white bracket. The posterior pharyngeal bulb is marked by a white arrowhead for orientation, and the anal depressor cell is marked by a black asterisk. C, transgenic animals expressing Pglr-1::dsRED and Pwdr-20::NLS::GFP were imaged. White arrowheads mark head neurons that express both reporters. D–F, representative images of the VNCs of L4 larval animals harboring a GLR-1::GFP transgene expressed under the control of the glr-1 promoter (nuIs25). The following genotypes are shown: wild type (D), wdr-20 (gk547140) (E), and wdr-20 (gk547140) expressing WDR-20 under the control of the glr-1 promoter (F, Rescue). The wdr-20 (gk547140) allele was backcrossed at least eight times prior to analysis. G, quantification of GLR-1::GFP punctum intensities (normalized, norm.) for the strains pictured in D--F. Shown are the means and S.E. for n = 30 (wild type), n = 27 (wdr-20 (gk547140)), and n = 27 (Rescue). H–J, representative images of the VNCs of L4 larval animals harboring a GLR-1::GFP transgene expressed under the control of the glr-1 promoter (nuIs25). The following genotypes are shown: wild type (H), wdr-48 (tm4575) (I), and wdr-48 (gk173034) (J). The wdr-48 (gk173034) allele was backcrossed at least six times prior to analysis. K, quantification of GLR-1::GFP punctum intensities (normalized) for the strains pictured in H–J. Shown are the means ± S.E. for n = 127 (wild type), n = 60 (wdr-48 (tm4575)), and n = 51 (wdr-48 (gk173034)). L, average number of reversals per minute for spontaneous locomotion assays performed on worms of the following genotypes: n = 28 (wild type), n = 16 (wdr-20 (gk547140)), n = 15 (wdr-48 (tm4575)), n = 13 (usp-46 (ok2232)). Values that differ significantly from the wild type (Tukey-Kramer test) are denoted above each bar. Other comparisons are marked by brackets. **, p < 0.001; n.s., no significant difference between the indicated strains (p ≥ 0.05). Error bars show S.E.