Fig. 5
SP-AH-mediated delivery of MIR376B into breast cancer cells blocked autophagy. a Quantitative PCR (qPCR) analysis of MIR376B levels in control (CNT, MIR376B only)-, SP-H plus MIR376B(SP-H/MIR376B)- or SP-AH plus MIR376B(SP-AH/MIR376B)-treated SKBR3 cells (mean ± SD of independent experiments, n = 5, **p < 0.03). a.u. arbitrary units. b qPCR analysis of BECN1 mRNA levels in SKBR3 cells treated as in (a) (n = 5, *p < 0.05). c qPCR analysis of ATG4C mRNA levels in SKBR3 cells treated as in (a) (n = 5, **p < 0.03). d Immunoblot analysis of BECN1 in the extracts from SP-AH- or SP-AH/MIR376B-treated SKBR3 cells. ACTB was used as the loading control. BECN1/ACTB band intensity ratios were marked. e Immunoblot analysis of ATG4C in the extracts from SP-AH- or SP-AH/MIR376B-treated SKBR3 cells. ACTB was used as loading control. ATG4C/ACTB band intensity ratios were marked. f qPCR analysis of MIR376B levels in CNT-, SP-H/MIR376B- or SP-AH/MIR376B-treated MDA cells (n = 5, ***p < 0.01). g qPCR analysis of BECN1 in MDA cells treated as in (f) (n = 5, *p < 0.05). h qPCR analysis of ATG4C in MDA cells treated as in (f) (n = 5, **p < 0.03). i Immunoblot analysis of BECN1 in the extracts from SP-AH- or SP-AH/MIR376B-treated MDA cells. j Immunoblot analysis of ATG4C in the extracts from SP-AH- or SP-AH/MIR376B-treated MDA cells. k Fluorescence microscopy images of SP-AH- or SP-AH/MIR376B-treated SKBR3 cells that were immunostained with an anti-LC3 antibody. l Graph showing the quantification of LC3 dot positivity in SKBR3 cells (n = 3, *p < 0.05). m Immunoblot analysis of LC3 in the extracts from SP-AH- or SP-AH/MIR376B-treated SKBR3 cells. LC3-II/I band intensity ratios were marked. n Fluorescence microscopy images of SP-AH- or SP-AH/MIR376B-treated MDA cells that were immunostained with an anti-LC3 antibody. o Graph showing the quantification of LC3 dot positivity in MDA cells (n = 3, **p < 0.03). p Immunoblot analysis of LC3 in the extracts from SP-AH- or SP-AH/MIR376B-treated MDA cells. LC3-II/I band intensity ratios were marked. Scale bar: 25 µm