Immunofluorescence Analysis of LC3 Expression

The three cell lines were transfected the miR-224-mimics/inhibitor as described above. Cells were fixed in 4% paraformaldehyde and permeabilized by 1% Triton X-100. Following blocking with 1% bovine serum albumin, cells were serially incubated in rabbit anti-LC3 (Abcam) and Goat anti-rabbit Alexa Fluo488 (Invitrogen). The images were acquired using the AV300-ASW confocal microscope (Olympus America Inc., Center Valley, USA) with a 60 × oil lens. The pictures analysed using Image-Pro Plus 6.0 (Media Cybernetics)32 (link).

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Fang W., Shu S., Yongmei L., Endong Z., Lirong Y, & Bei S. (2016). miR-224-3p inhibits autophagy in cervical cancer cells by targeting FIP200. Scientific Reports, 6, 33229.

Publication 2016

rabbit anti-LC3 Goat anti-rabbit Alexa Fluo488 AV300-ASW confocal microscope Image-Pro Plus 6.0

Bovine serum albumin Cell lines Cells Confocal microscope Goat Lens Paraformaldehyde Rabbit Triton x 100

Corresponding Organization :

Other organizations : Tianjin Medical University, Second Hospital of Tianjin Medical University

Top 5 similar protocols

Variable analysis

independent variables

MiR-224-mimics
MiR-224-inhibitor

dependent variables

LC3 expression

control variables

Cell lines
Paraformaldehyde fixation
Triton X-100 permeabilization
Bovine serum albumin blocking
Primary antibody (rabbit anti-LC3)
Secondary antibody (Goat anti-rabbit Alexa Fluo488)
Confocal microscope (Olympus AV300-ASW)

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