Figure 4.

Decreased protein synthesis and ribosome large subunits in the absence of RPL39L

(A) Confocal images of testis cross-sections immunostained with anti-puromycin following in vivo labeling. Scale bar: 100 μm.

(B) Puromycin⁺ cells per cross-section as shown in (A) N ≥ 41, One-way ANOVA Tukey’s test, ∗∗∗p < 0.001.

(C) Confocal images of dispersed testicular cells immunostained with anti-puromycin following in vitro labeling. Scale bar: 50 μm.

(D) Western blotting of testis lysates following in vivo puromycin labeling. Shown are two experimental repeats.

(E) Western blotting of testicular cell lysates following in vitro puromycin labeling. α-TUBULIN was used as loading control in (D) and (E).

(F and G) Confocal images of testis sections immunostained with anti-puromycin and anti-UCHL1 (F) or anti-SOX9 (G) following in vivo puromycin labeling. Cell nuclei were stained with DAPI. Scale bars: 50 μm.

(H) Sucrose gradient sedimentation of testis lysates. Peaks of 40S SSU, 60S LSU and 80S monosomes are indicated with blue boxes.

(I and J) GO analyses of proteins pulled-down by both GST-RPL39 and GST-RPL39L. GO groups relevant to ribosome biogenesis are indicated in green.

(K) Co-immunoprecipitation of testis lysates with pan-anti-RPL39 or anti-NOP10. NOP10 is common to both GST-RPL39 and GST-RPL39L pull-downs.