Increasing emb-9 expression decreases the LET-2/EMB-9 BM ratio. (A) Top: a schematic diagram of a day 1 adult showing the head region and the location of the body wall muscle BM analyzed (yellow box). Below: LET-2::mNG (C) with and without emb-9::mRuby2 overexpression and EMB-9::mNG (C) with and without emb-9::mNG::mRuby2 overexpression in the body wall muscles of day 1 adults. The yellow arrows indicate body wall muscle BM and yellow arrowheads the intracellular aggregates. Aggregates were more prevalent in EMB-9::mNG (C) animals after emb-9 overexpression (see Fig. S5 F). The scale bar is 5 µm. (B) Top: boxplots show percent gain of LET-2::mNG (C) and EMB-9::mNG (C) protein (mean fluorescence intensity) in the body wall muscle BM after emb-9 overexpression. For each trial, mean fluorescence intensity measurements of LET-2::mNG (C) and EMB-9::mNG (C) with emb-9 overexpression were normalized to the average of all mean fluorescence intensity measurements of LET-2::mNG (C) and EMB-9::mNG (C) without overexpression (see Materials and methods, n > 5 animals for each of three trials. ****P > 0.0001, unpaired two-tailed Student’s t test with Welch’s correction). Bottom: the percent gain of both proteins for each trial is listed in the table. Colors in the table correspond to data points in the boxplot. (C) Top: boxplots show the LET-2::mNG (C)/EMB-9::mNG (C) mean fluorescence ratios in the body wall muscle BM with and without emb-9 overexpression. LET-2::mNG (C) mean fluorescence intensity measurements were normalized to the average of all EMB-9::mNG (C) mean fluorescence intensity measurements within each trial (n > 5 animals for each of the three trials. ****P > 0.0001, unpaired two-tailed Student’s t test with Welch’s correction). Bottom: table showing the ratios of the average BM fluorescence levels of LET-2::mNG (C)/EMB-9::mNG (C) for each trial. Colors in the table correspond to data points in the boxplot.