FIGURE SUMMARY
Title

Pregnancy-associated plasma protein-aa supports hair cell survival by regulating mitochondrial function

Authors
Alassaf, M., Daykin, E.C., Mathiaparanam, J., Wolman, M.A.
Source
Full text @ Elife
Figure 1—source data 2.

Mean F(TMRE) and ratio of mean F(TMRE) to mean F(GFP) in wild type and <italic>pappaa<sup>p170</sup></italic> hair cells following treatment with NVP-AEW541.

Figure 2.

<italic>pappaa</italic> is expressed by neuromast support cells and motor neurons.

(A) Whole mount in situ hybridization shows pappaa mRNA expression at four dpf by lateral line neuromasts (arrowheads). (B) Fluorescent in situ hybridization of pappaa (green) and brn3c:GFP labeled hair cells (white) shows pappaa mRNA expression by the support cells that surround hair cells. (C) RT-PCR of fluorescently sorted brn3c:GFP labeled hair cells and mnx1:GFP labeled motor neurons shows pappaa expression by motor neurons, but not hair cells. RT-PCR products represent pappaa cDNA fragment.
Figure 3—figure supplement 1—source data 1.

Support cells are not affected by neomycin treatment.

(A) Mean percentage of surviving neuromast support cells at 4 hr post-neomycin treatment. To calculate support cell survival percentage, support cell number 4 hr post-neomycin treatment was normalized to mean support cell number in vehicle treated larvae in the same genotype. Two-way ANOVA, Holm-Sidak post test revealed no significant difference among groups. N = 6–9 larvae per group (shown at base of bars). Total number of neuromasts included in the analysis = 27 (wild type; vehicle-treated), 18 (wild type; 1 µM neomycin), 21 (wild type; 10 µM neomycin), 24 (wild type; 30 µM neomycin), 27 (pappaap170; vehicle-treated), 18 (pappaap170; 1 µM neomycin), 18 (pappaap170; 10 µM neomycin), and 18 (pappaap170; 30 µM neomycin). Error bars = SEM. (B) Representative confocal images of anti-SOX2 labeled support cells that were control or 30 µM neomycin treated. Scale = 10 µm.

10.7554/eLife.47061.008

Support cell survival post neomycin in wild type and <italic>pappaa<sup>p170</sup></italic>larvae.

Figure 3—figure supplement 1—source data 1.

Support cells are not affected by neomycin treatment.

(A) Mean percentage of surviving neuromast support cells at 4 hr post-neomycin treatment. To calculate support cell survival percentage, support cell number 4 hr post-neomycin treatment was normalized to mean support cell number in vehicle treated larvae in the same genotype. Two-way ANOVA, Holm-Sidak post test revealed no significant difference among groups. N = 6–9 larvae per group (shown at base of bars). Total number of neuromasts included in the analysis = 27 (wild type; vehicle-treated), 18 (wild type; 1 µM neomycin), 21 (wild type; 10 µM neomycin), 24 (wild type; 30 µM neomycin), 27 (pappaap170; vehicle-treated), 18 (pappaap170; 1 µM neomycin), 18 (pappaap170; 10 µM neomycin), and 18 (pappaap170; 30 µM neomycin). Error bars = SEM. (B) Representative confocal images of anti-SOX2 labeled support cells that were control or 30 µM neomycin treated. Scale = 10 µm.

10.7554/eLife.47061.008

Support cell survival post neomycin in wild type and <italic>pappaa<sup>p170</sup></italic>larvae.

Figure 4—source data 2.

Hair cell survival post neomycin in wild type and <italic>Tg(hsp70:dnIGF1Ra-GFP)</italic> larvae.

Figure 5—figure supplement 1—source data 2.

Mean F(CellROX) and ratio of mean F(CellROX) to mean F(GFP) in wild type and pappaa mutant hair cells.

Figure 5—figure supplement 1—source data 2.

Mean F(CellROX) and ratio of mean F(CellROX) to mean F(GFP) in wild type and pappaa mutant hair cells.

Figure 6—source data 3.

Hair cell survival post Cyclosporin A in wild type and <italic>pappaa<sup>p170</sup></italic> larvae.

Acknowledgments
This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ Elife
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