1. Variance in total levels of phospholipase C zeta (PLC-ζ) in human sperm may limit the applicability of quantitative immunofluorescent analysis as a diagnostic indicator of oocyte activation capability 
Junaid Kashir, Ph.D., Celine Jones, Ginny Mounce, M.Sc., Walaa M. Ramadan, M.Sc., Bernadette Lemmon, B.M., Bjorn Heindryckx, Ph.D., Petra de Sutter, M.D., Ph.D., John Parrington, Ph.D., Karen Turner, Ph.D., Tim Child, M.D., Enda McVeigh, M.D., Kevin Coward, Ph.D. 
Fertility and Sterility 
Volume 99, Issue 1, Pages e3 (January 2013)
DOI: /j.fertnstert
Copyright © 2013 American Society for Reproductive Medicine Terms and Conditions

2. Figure 1
Mean relative total phospholipase C zeta (PLC-ζ) fluorescence levels exhibited by sperm from individual control and oocyte activation–deficient (OAD) patients. Fluorescence intensity was quantified in arbitrary units using ImageJ software. Asterisks (*; **; ***) denote statistically significant differences (P≤.05), whereas hash marks (#; ##; ###; #′; ##′) denote statistically insignificant differences. Combinations of marks indicate comparisons between different groups (e.g., # indicates comparison with one group, and ## indicates comparison with another). Data are shown as mean ± SEM.
Fertility and Sterility  , e3DOI: ( /j.fertnstert )
Copyright © 2013 American Society for Reproductive Medicine Terms and Conditions

3. Figure 2
(A) Representative confocal images of phospholipase C zeta (PLC-ζ) immunofluorescence in human sperm showing characteristic localization patterns previously identified by Grasa et al. (2008). However, additional patterns, not previously described by Grasa et al., were identified in the present study. Images were captured at ×63 optical magnification and represent overlays of PLC-ζ (green), and differential interference contrast images. White scale bars indicate 5 μm. White arrow indicates acrosomal localization; black arrow indicates equatorial localization; and red arrows indicate postacrosomal localization. The rightmost panel indicates sperm exhibiting no PLC-ζ immunoreactivity. (B) Mean proportions of sperm (percentages) exhibiting PLC-ζ immunofluorescence from control subjects and oocyte activation–deficient (OAD) patients. Proportions were determined with the use of ImageJ software. The asterisks (*; **) on the histogram denote statistically significant differences (P≤.05), whereas hash marks (#; ##; ###) denote statistically insignificant differences (P≥.05). Combinations of marks indicate comparisons between different groups (e.g., * indicates comparison with one group, and ** indicates comparison with another). Data are shown as mean ± SEM.
Fertility and Sterility  , e3DOI: ( /j.fertnstert )
Copyright © 2013 American Society for Reproductive Medicine Terms and Conditions

4. Supplemental Figure 1
Mean relative total phospholipase C zeta (PLC-ζ) fluorescence levels exhibited by control and oocyte activation–deficient (OAD) patient sperm. Fluorescence intensity was quantified in arbitrary units with the use of ImageJ software. Asterisk (*) denotes a statistically significant difference (P≤.05). Data are shown as mean ± SEM.
Fertility and Sterility  , e3DOI: ( /j.fertnstert )
Copyright © 2013 American Society for Reproductive Medicine Terms and Conditions

5. Supplemental Figure 2
Mean proportions of sperm (percentages) exhibiting phospholipase C zeta (PLC-ζ) immunofluorescence from control and oocyte activation–deficient (OAD) sperm. Proportions were determined with the use of ImageJ software. Asterisk (*) denotes a statistically significant difference (P≤.05). Data are shown as mean ± SEM.
Fertility and Sterility  , e3DOI: ( /j.fertnstert )
Copyright © 2013 American Society for Reproductive Medicine Terms and Conditions

6. Supplemental Figure 3
Mean proportions of sperm (percentages) exhibiting phospholipase C zeta (PLC-ζ) immunofluorescence patterns in total sperm from (A) control subjects and (B) oocyte activation–deficient (OAD) patients. Proportions were determined with the use of ImageJ software. (A) The asterisk (*) denotes statistically significant differences (P≤.05), whereas hash marks (#; #′; ##; ###) on the histogram denote statistically insignificant differences (P≥.05). (B) Asterisks (*; **; **′; ***) on the histogram denote statistically significant differences (P≤.05). Combinations of asterisks and hash marks on both histograms indicate comparisons between different groups (e.g., # indicates comparison with one group, and ## indicates comparison with another). Data are shown as mean ± SEM.
Fertility and Sterility  , e3DOI: ( /j.fertnstert )
Copyright © 2013 American Society for Reproductive Medicine Terms and Conditions