Appendix A. Calculations to demonstrate that the major contributor of Coronin-1A in BCa patient urine is not from lysed WBCs but from the bladder tumor.

Albumin determination
Concentration of albumin in blood*	40 μg/μl
Concentration range of albumin in BCa patient urine samples&	0.012-2.56 μg/μl
Mean albumin concentration in BCa patient urine samples	1.18 μg/μl
Approximate volume of blood in urine#	(1/40)×1.18 = 0.0295 μl
White Blood Cells (WBC) determination
No. of WBC/μl of blood*	~5000 WBCs
Hence, ~0.0295 μl of blood in urine contains	(5000×0.0295) =147.5 WBCs
Coronin-1A calculation
Weight of a single cell*	3.5×10−3μg
Amount of protein/cell§,*	0.0007 μg
40 μg of lysed WBC from buffy coat would be derived from	(1/0.0007)×40 ≈ 57,000 WBCs¥
Coronin-1A blot intensity of 57,000 WBC lysate£	34,403
Hence, 147.5 WBCs if lysed in urine would produce intensity of	(34,403/57000)×147.5 = 89.025
Actual mean Coronin-1A intensity from BCa urine sample blot	25,788.58¶
Fold-change in intensity for Coronin-1A	(25,788.58/89.025) = 289.68

^*Reported in the literature

^&Deduced from our experimental analysis; further calculations were done using the higher limit of the range.

^#Assumption: Albumin source is solely blood

^§Protein weight is 20% of cells weight

^¥Rounded down for calculations. Actual calculation is 57,142.86 WBCs

^£Result obtained from western blotting experiments conducted in our lab

^¶Averaged value.