Quantification of Renal Cell Markers

The procedure and protocols have been described by previous reports [8 (link),16 (link),29 (link),30 (link)]. Briefly, sections were incubated with primary antibodies specifically against CD31 (1:200, BD Pharmingen, Franklin Lakes, NJ, USA), vWF (1:200, Abcam), CXCR4 (1:200, Abcam) and SDF-1α (1:100, Santa Cruz), while sections incubated with irrelevant antibodies served as controls. Three sections of kidney specimens from each rat were analyzed. For quantification, three random HPFs (400× for IHC and IF studies) were analyzed in each section. The mean number (expressed as a percentage) of positively-stained cells per HPF for each animal was first divided by the total positively DAPI-stained cells, then determined by the summation of all numbers divided by 9.

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Chen Y.C., Sheu J.J., Chiang J.Y., Shao P.L., Wu S.C., Sung P.H., Li Y.C., Chen Y.L., Huang T.H., Chen K.H, & Yip H.K. (2020). Circulatory Rejuvenated EPCs Derived from PAOD Patients Treated by CD34+ Cells and Hyperbaric Oxygen Therapy Salvaged the Nude Mouse Limb against Critical Ischemia. International Journal of Molecular Sciences, 21(21), 7887.

Publication 2020

CXCR4 SDF-1α

Animal Antibodies Cells Cxcr4 Dapi Kidney

Corresponding Organization : Xiamen Chang Gung Hospital

Other organizations : Kaohsiung Medical University, Ministry of Health and Welfare, National Sun Yat-sen University

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Variable analysis

independent variables

Primary antibodies specifically against CD31, vWF, CXCR4, and SDF-1α

dependent variables

Mean number (expressed as a percentage) of positively-stained cells per high-power field (HPF) for each animal

control variables

Sections incubated with irrelevant antibodies

controls

Positive control: Sections incubated with primary antibodies specifically against CD31, vWF, CXCR4, and SDF-1α
Negative control: Sections incubated with irrelevant antibodies

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