miR-29c-3p Expression Analysis by FISH

FISH was performed using the previously described protocol (Shi et al., 2018 (link); Moldovan et al., 2019 (link)). The digoxigenin-labeled miR-29c-3p was hybridized with the disturbed LNA probe (Exiqon) at 61°C. The signal in situ was stained with digoxin-labeled anti-digoxin AP antibody (Roche Diagnostics GmbH, Mannheim, Germany), and developed with BM purple substrate (Roche). Fluorescence intensity was quantified by the Image Pro Plus 6.0 software (Media Cybernetics).

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Lin B., Zhu J., Yin G., Liao M., Lin G., Yan Y., Huang D, & Lu S. (2021). Transcription Factor DLX5 Promotes Hair Follicle Stem Cell Differentiation by Regulating the c-MYC/microRNA-29c-3p/NSD1 Axis. Frontiers in Cell and Developmental Biology, 9, 554831.

Publication 2021

BM purple substrate Image Pro Plus 6.0 software

Anti antibody Diagnostics Digoxigenin Digoxin Fish Fluorescence

Corresponding Organization : Guangxi Medical University

Other organizations : Guangxi Academy of Sciences

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Variable analysis

independent variables

MiR-29c-3p probe hybridization temperature (61°C)

dependent variables

Fluorescence intensity

control variables

Previously described FISH protocol (Shi et al., 2018; Moldovan et al., 2019)
Digoxigenin-labeled miR-29c-3p probe
Disturbed LNA probe (Exiqon)
Digoxin-labeled anti-digoxin AP antibody (Roche Diagnostics GmbH, Mannheim, Germany)
BM purple substrate (Roche)

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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