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Fig. 5 | Journal of Experimental & Clinical Cancer Research

Fig. 5

From: Knockdown of HMGA2 regulates the level of autophagy via interactions between MSI2 and Beclin1 to inhibit NF1-associated malignant peripheral nerve sheath tumour growth

Fig. 5

HMGA2 regulates autophagy via interactions between MSI2 and Beclin1 to regulate the growth of NF1 MPNSTs. a MSI2 protein expression is elevated in MPNSTs compared with that in neurofibromas. b GAPDH was used as the control. Relative MSI2 protein level was expressed as a percentage of GAPDH. c IHC for positive and negative MSI2 staining in normal human nerve, neurofibroma, and NF1 MPNST samples compared to sporadic MPNST samples. Scale bar, 50 μm. d Overall survival of MPNST patients with positive MSI2 staining. e Correlation analysis between positive HMGA2 and MSI2 staining in 57 MPNST tissues. f and g EdU (red) assays for proliferation rates. Nuclei are stained with Hoechst 33342 (blue). Scale bar = 50 μm. Overexpression of MSI2 reverses proliferation activity of shHMGA2 cells. h and i TUNEL positivity in HMGA2 knockdown cells was markedly decreased upon MSI2 overexpression. Scale bar = 50 μm. j Percentage of apoptotic cells was determined by FCM. Overexpression of MSI2 reverses the apoptosis level of shHMGA2 cells. k Cells co-transfected with shHMGA2 and MSI2 showed LC3-II fluorescence puncta with increased LC3-II levels compared to those in cells transfected with shHMGA2 alone. l WB was performed to detect the indicated proteins in control-, shHMGA2- and shHMGA2 + MSI2-transfected NF1 MPNST cells. m and n qRT-PCR analysis was performed to detect HMGA2, MSI2, PTEN, and Beclin1 mRNA expression in control-, shHMGA2- and shHMGA2 + MSI2-transfected NF1 MPNST cells. Bars represent the SEM. **P < 0.05 by Student’s t-tests. o IP and WB verifying interactions between MSI2 and Beclin1 in NF1 MPNST cells. p Immunofluorescence analysis of the co-localization and expression of MSI2 and Beclin1 in NF1 MPNST cells

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