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. 2021 Jul 18;42(4):514–524. doi: 10.24272/j.issn.2095-8137.2021.041

Figure 3.

Figure 3

Heat stress induces spermatocyte apoptosis and endoplasmic reticulum stress

A: mRNA expression of heat shock genes (Hsp27 and Hsp70) in spermatocytes (GC2 cell line) at 0, 2, 4, and 6 h after heat treatment for 2 h. B: mRNA expression of apoptosis genes (Caspase3 and Chop) and Grp78 in spermatocytes at 0, 2, 4, and 6 h after heat treatment for 2 h. C: mRNA expression of ERS-related genes (Ire1, Perk and Atf6) in spermatocytes at 0, 2, 4, and 6 h after heat treatment for 2 h. D: Western blotting of ERS-related proteins (p-IRE1, p-PERK, and ATF6), apoptosis protein (CASPASE12), and heat shock protein (HSP70) in spermatocytes at 0, 2, 4, and 6 h after heat treatment for 2 h. E: Image J analysis of (D). F: TUNEL staining for apoptosis at 4 h after heat treatment for 2 h. Cells showing red fluorescence are positive. Hoechst 33342 (blue)-stained cell nuclei. G: Immunofluorescent images showing GRP78 and CHOP in spermatocytes at 4 h after heat treatment for 2 h. H: Immunofluorescent images showing heat shock proteins (HSP27 and HSP70) in spermatocytes at 4 h after heat treatment for 2 h. I: Immunofluorescent images showing ERS-related proteins (p-IRE1, p-PERK, and ATF6) in spermatocytes at 4 h after heat treatment for 2 h. All images were merged from pIRE1, pPERK, or ATF6 (green) and Hoechst 33342-stained nuclei (blue). Scale bar: 200 μm. Con: Control, hs: Heat stress. Data are mean±SD (n=3); statistical significance is expressed as: **: P<0.01, NS: Not significant.