Fig. 7: Loss of BRA-2 restores nuclear clustering and PLK-2 loading in htp-1 mutants.
a Nuclei in transition zone (TZ) or mid-pachytene (MP) stained for PLK-2 (yellow)/SYP-1 (magenta) counterstained by DAPI (cyan) in HA::AID::bra-2; TIR1::mRuby control animals and htp-1(gk174) mutants, before and after auxin exposure (24 h). Scale bar 5 μm. Analysis was performed in biological duplicates. b Mid-pachytene (MP) nuclei stained for HTP-3 (red)/SYP-1 (green) in the indicated genetic backgrounds before and after exposure to auxin. Scale bar 5 μm. Analysis was performed in biological duplicates. c Quantification of nuclei with paired signals for Ch. V in the indicated genetic backgrounds before and after exposure to auxin. Bars depict S.E.M. and asterisks indicate statistical significance as assessed by Χ2 test (two-sided, CI. = 0.05, *=p < 0.0001, ns= non- significant). At least three germ lines for each genetic background were employed for quantification, and the number of nuclei scored was: HA::AID::bra-2 (-auxin, zone 1-5) 150, 179, 175, 146, 110; HA::AID::bra-2; htp-1 (-auxin, zone 1-5) 128, 139, 148, 174, 122; HA::AID::bra-2 (+auxin, zone 1-5) 198, 208, 200, 200, 126; HA::AID::bra-2; htp-1 (+auxin, zone 1-5) 155, 174, 175, 163, 116. d Quantification of non-homologous synapsis in the indicated genetic backgrounds and exposure conditions to auxin only in the pachytene region. Quantifications were performed on the same samples quantified in (c), but only in nuclei spanning the pachytene region and corresponding to zones 4 (EP/MP) and 5 (MP/LP). Bars indicate S.E.M. and asterisk denotes statistical significance assessed by Χ2 test (two-sided, CI. = 0.05, ns= non-significant, *p < 0.0001). e Representative images of mid-pachytene nuclei showing FISH for Ch. V (orange) and anti-SYP-1 (light green) immunostaining, counterstained by DAPI (purple). Scale bar 5 μm. Analysis was performed in biological triplicates.
