Fig. 2: HSF5 expression pattern in the mouse seminiferous tubules.
a Seminiferous tubule sections in WT testis (8 weeks old) were immunostained as indicated. pL preleptotene, L Leptotene, Z Zygotene, ePa early Pachytene, P Pachytene, M I Metaphase I, int Interkinesis, rS round Spermatid, eS elongated Spermatid. Boundaries of the seminiferous tubules are indicated by white dashed lines. * indicates a non-specific cross-reactivity of the gunia pig anti-SYCP3 antibody to sperm tail. HSF5 was immunostained by HSF5-C antibody. The same immunostaining pattern of HSF5 was confirmed by other HSF5-N1 and HSF5-N2 antibodies, as shown in Fig. S2a. Roman numbers indicate the seminiferous tubule stages. Biologically independent mice (n = 3) were examined in three separate experiments. Scale bar: 25 μm. b The schematic of the Hsf5-3xFLAG-HA knockin allele. c Testis extracts from Hsf5-3xFLAG-HA knockin and negative control WT mouse testis (5 weeks old) were immunoblotted as indicated. Red arrow indicates HSF5-3xFLAG-HA protein derived from the knockin allele. Blue arrow indicates HSF5 protein derived from WT allele. d Seminiferous tubule sections in Hsf5-3xFLAG-HA knockin (n = 1) and negative control WT mouse testis (n = 1) at 5 weeks old were immunostained as indicated. Scale bar: 25 μm. e Seminiferous tubule sections in WT testis (P16) were immunostained as indicated. Arrow and arrowhead indicate HSF5-positive/H1t-positive and HSF5-negative/H1t-positive pachytene spermatocytes, respectively. Scale bar: 25 μm. A single experiment was performed. f The schematic of expression of HSF5 (green), H1t (blue), HSF1(red)46, in the stages of the seminiferous tubules.
