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. Author manuscript; available in PMC: 2013 Apr 1.
Published in final edited form as: J Immunol. 2012 Feb 27;188(7):3268–3277. doi: 10.4049/jimmunol.1103368

Figure 6. Kidney deposition and urinary excretion of mC5b-9.

Figure 6

A. Indirect IF staining shows granular GBM deposition of mC5b-9 in α3NC1-immunized wild type (a) and FcRγ−/− (b) mice, but not in adjuvant-immunized mice (c). Staining was negative when the primary Ab was omitted (d). B. Western blot analysis of mouse urine samples (5 μl/lane) separated by SDS-PAGE under non-reducing conditions using Abs to mC5b-9 showed specific staining of a ~250 kDa band (arrow) in urine collected at 8 weeks (lane f), but not at 0, 2, 4, or 6 weeks after immunization with α3NC1 (lanes b–e, respectively), nor in urine collected at 8 weeks after immunization with CFA (lane a). Urine from a 7-months old B6.Col4a3−/− mouse with heavy albuminuria (ACR = 103) was used as an additional control. The band at ~150 kDa was also stained when the primary Ab was omitted, suggesting it is due to secondary Ab cross-reacting with excreted mIgG in mouse urine.