Immunohistochemical Evaluation of Stem Cell Markers

The procedure of IHC and ICC staining was shown in the previous studies [52 (link)]. LGR6 staining was evaluated blindly and independently by two pathologists. The score for staining intensity (0 = no staining, 1 = light brown, 2 = brown, and 3 = dark brown) and staining frequency (0 = 0–10%, 1 = 11–25%, 2 = 26–50%, 3 = 51–75%, and 4 = 76–100%) were multiplied to obtain the immunoreactivity score (IRS). The IRS 0–3 was deemed negative, 4–6 weak positive, and >6 strong positive. Two different pathologists evaluated all the specimens in a blinded manner. The antibodies used were as follows: anti-LGR6 (#MAB8458, R&D Systems, USA), anti-β-catenin (#sc-7963, Santa Cruz, USA), anti-TCF7L2 (#sc-166699, Santa Cruz, USA), anti-c-Myc (#10828-1-AP, Proteintech, China), OCT4 (#sc-5279, Santa Cruz, USA), anti-SOX2 (#3579, Cell Signaling Technology, USA), anti-KLF4 (#sc-20691, Santa Cruz, USA), anti-ALDH1A1 (#sc-374149, Santa Cruz, USA), anti-LGR5 (#PAB2591, Abnova, Taiwan), anti-LGR4 (#sc-390630, Santa Cruz, USA).

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Feng Q., Li S., Ma H.M., Yang W.T, & Zheng P.S. (2021). LGR6 activates the Wnt/β-catenin signaling pathway and forms a β-catenin/TCF7L2/LGR6 feedback loop in LGR6high cervical cancer stem cells. Oncogene, 40(42), 6103-6114.

Publication 2021

anti-β-catenin anti-c-Myc anti-SOX2 anti-KLF4 anti-ALDH1A1

Aldh1a1 Antibodies Klf4 Light Oct4 Pathologists Sox2 Tcf7l2 Weak β catenin

Corresponding Organization :

Other organizations : First Affiliated Hospital of Xi'an Jiaotong University, Ministry of Education of the People's Republic of China

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Variable analysis

independent variables

Staining intensity (0 = no staining, 1 = light brown, 2 = brown, and 3 = dark brown)
Staining frequency (0 = 0–10%, 1 = 11–25%, 2 = 26–50%, 3 = 51–75%, and 4 = 76–100%)

dependent variables

Immunoreactivity score (IRS)

control variables

The procedure of IHC and ICC staining was kept constant as described in the previous studies [52]
Two different pathologists evaluated all the specimens in a blinded manner

positive controls

Not explicitly mentioned

negative controls

Not explicitly mentioned

Annotations

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