Journal: The Journal of Biological Chemistry

Article Title: The ER-associated degradation adaptor SEL1L is dispensable for ER homeostasis and the differentiation of spermatogenic cells

doi: 10.1016/j.jbc.2025.110283

Figure Lengend Snippet: SEL1L and HRD1 are expressed in spermatogenic cells. A and B , Western blot analysis of various tissues from 6-week-old wild-type mice, with quantitation of SEL1L and HRD1 shown in ( B ). With equal protein loading across samples, SEL1L and HRD1 quantification was not normalized due to the varying expression of housekeeping proteins β-actin and HSP90. N = 3. Values, mean ± SD. ∗∗, p < 0.01; ∗∗∗, p < 0.001; ns, not significant comparing different tissues to the liver by one-way ANOVA with Tukey multiple comparison test. C and D , Western blot analysis of testes from wild-type mice at different ages, with quantitation normalized to HSP90 shown in ( D ). N = 3. Values, mean ± SD. ∗, p < 0.05; ∗∗, p < 0.01; ∗∗∗, p < 0.001 comparing different ages to 7 days by one-way ANOVA with Tukey multiple comparison test. E – G , immunofluorescent staining of testes from wild-type mice at 7, 14, and 35 days old ( E ) and 6 weeks old ( F and G ), showing the expression of SEL1L and HRD1 in various cell types as illustrated. GCNA labels spermatogonia; lectin PNA labels acrosomes in round and elongated spermatids. In ( E ), the average immunofluorescent signal intensity of SEL1L and HRD1 in individual seminiferous tubules was quantitated. 60 to 75 seminiferous tubules from three individual mice were quantitated at each age. Values, mean ± SD. ∗∗∗, p < 0.001 by one-way ANOVA with Tukey multiple comparison test. In ( F and G ), insets of higher magnification are shown below. Representative data from n = 3 in ( E ) and n = 4 in ( F and G ).

Article Snippet: The following antibodies were used in this study: SEL1L (Abcam ab78298), HRD1 (Abclonal customized E15102 ), GRP94 (Novus NBP2-42379), DDX4 (Abcam ab13840), lectin PNA (Invitrogen L21409 ), GCNA (DSHB 10D9G11), and vimentin (Biolegend 699301).

Techniques: Western Blot, Quantitation Assay, Expressing, Comparison, Staining

Journal: The Journal of Biological Chemistry

Article Title: The ER-associated degradation adaptor SEL1L is dispensable for ER homeostasis and the differentiation of spermatogenic cells

doi: 10.1016/j.jbc.2025.110283

Figure Lengend Snippet: Generation of male germ cell-specific Sel1L knockout mice. A , diagram illustrating the generation of male germ cell-specific Sel1L knockout mice ( KO Stra8 ), wildtype ( WT Stra8 ), and heterozygous ( HET Stra8 ) littermates. B , diagram of the Sel1L locus with primers for genotyping illustrated. C , representative genotyping results. D , Western blot analysis of testes from 6-week-old WT Stra8 , HET Stra8 , and KO Stra8 littermates, with quantitation of SEL1L normalized to HSP90 shown below. N = 4 per cohort. E – H , immunofluorescent staining of testes from 6-week-old WT Stra8 , HET Stra8 , and KO Stra8 littermates, showing the ablation of SEL1L in spermatocytes and spermatids ( E ), but not in GCNA + spermatogonia ( G ), or vimentin + Sertoli cells ( H ). Insets of higher magnification are shown on the right in ( E ). The immunofluorescent signal intensity of SEL1L in individual seminiferous tubules is quantified in ( F ). In ( H ), ∗ indicates non-specific staining by the SEL1L antibody in spermatid flagella at stage VII-VIII. Values, mean ± SD. ∗, p < 0.05; ∗∗∗, p < 0.001 by one-way ANOVA with Tukey multiple comparison tests. Representative data from n = 4 mice per cohort.

Article Snippet: The following antibodies were used in this study: SEL1L (Abcam ab78298), HRD1 (Abclonal customized E15102 ), GRP94 (Novus NBP2-42379), DDX4 (Abcam ab13840), lectin PNA (Invitrogen L21409 ), GCNA (DSHB 10D9G11), and vimentin (Biolegend 699301).

Techniques: Knock-Out, Western Blot, Quantitation Assay, Staining, Comparison

Journal: The Journal of Biological Chemistry

Article Title: The ER-associated degradation adaptor SEL1L is dispensable for ER homeostasis and the differentiation of spermatogenic cells

doi: 10.1016/j.jbc.2025.110283

Figure Lengend Snippet: Sel1L ablation leads to the loss of HRD1 in spermatids, but not spermatocytes. A , Western blot analysis of testes from 6-week-old WT Stra8 , HET Stra8 , and KO Stra8 littermates, with quantitation of SEL1L and HRD1 normalized to HSP90 shown below. WT and SEL1L −/− HEK293T cells were shown as controls. B – D , immunofluorescent staining of testes from 6-week-old WT Stra8 and KO Stra8 littermates, showing HRD1 expression in spermatocytes and its absence in round and elongated spermatids. Lectin PNA labels acrosomes in round and elongated spermatids; GCNA labels spermatogonia. Representative data from n = 4 mice per cohort. E , Western blot analysis of ERAD substrate proteins in testes from 6-week-old WT Stra8 , HET Stra8 , and KO Stra8 littermates, with quantitation normalized to HSP90 shown below. Note the accumulation of ERAD substrates in SEL1L −/− HEK293T cells but not KO Stra8 testes. In ( A and E ), N = 4 for testes, n = 3 for HEK293T cells. Values, mean ± SD. ∗, p < 0.05; ∗∗, p < 0.01; ∗∗∗, p < 0.001 by one-way ANOVA with Tukey multiple comparison tests.

Article Snippet: The following antibodies were used in this study: SEL1L (Abcam ab78298), HRD1 (Abclonal customized E15102 ), GRP94 (Novus NBP2-42379), DDX4 (Abcam ab13840), lectin PNA (Invitrogen L21409 ), GCNA (DSHB 10D9G11), and vimentin (Biolegend 699301).

Techniques: Western Blot, Quantitation Assay, Staining, Expressing, Comparison

Journal: The Journal of Biological Chemistry

Article Title: The ER-associated degradation adaptor SEL1L is dispensable for ER homeostasis and the differentiation of spermatogenic cells

doi: 10.1016/j.jbc.2025.110283

Figure Lengend Snippet: Spermatogenesis is not affected by male germ cell-specific Sel1L ablation. Immunofluorescent staining of testes from 6-week-old WT Stra8 and KO Stra8 littermates, showing ( A ) GCNA, a germ-cell specific marker. B , DDX4, a spermatocyte and round spermatid-specific marker. C , lectin PNA, a sperm acrosome marker at different stages of the seminiferous epithelium cycle. Insets of higher magnification shown on the right . Representative images from n = 4 mice per cohort.

Article Snippet: The following antibodies were used in this study: SEL1L (Abcam ab78298), HRD1 (Abclonal customized E15102 ), GRP94 (Novus NBP2-42379), DDX4 (Abcam ab13840), lectin PNA (Invitrogen L21409 ), GCNA (DSHB 10D9G11), and vimentin (Biolegend 699301).

Techniques: Staining, Marker