Fig 1: Silencing efficiency of ITGB4 and lung structure of ITGB4f/f and ITGB4ccsp.cre mice. (A) ITGB4 expression was detected by conducting immunofluorescence. CCSP (red) and ITGB4 (green) were performed in lung sections. DAPI was used to stain cell nuclei (blue).Scale bars: 50 µm (B) Dilated airspaces with thickened alveolar septa were seen in H&E-stained paraffin sections of P28 lungs. (Scale bars: 250 µm for left and 50 µm for right) (C,D) RT-PCR analysis of ITGB4, integrin alpha6, integrin beta1 mRNA expression levels in ITGB4f/fand ITGB4ccsp.cre mice. (**p < 0.01). Data are presented as mean ± SD.
Fig 2: Normal and stiff substrate models were constructed in vitro. (A,B) Detection of contact angle and the contact angle before and after plasma treatment (C,D) ITGB4 expression of 16HBE14o-cells on the normal and stiff substrate by RT-PCR and western blot. (n = 3 independent experiments, *p < 0.05, **p < 0.01). Data are presented as mean ± SD.
Fig 3: PSMA promotes GBM development in vivo. (A) Bioluminescence imaging was performed to evaluate the growth of xenografts, (B) and the intensity of which was scanned as a representation of tumor growth. (C,D) The expression of PSMA, ITGB4, and phosphorylation of NF-?B p65 were also detected after using inhibitor 2-PMPA in vivo. Data were presented as mean with SD. *P < 0.05.
Fig 4: PSMA promotes GBM angiogenesis through interacting with ITGB4. (A) HUVECs with or without PSMA overexpression were lysed and immunoprecipitated with anti-PSMA antibody. (B) The expression of PSMA and ITGB4 was detected in input complex, and the expression of PSMA was detected in IP complex. (C) The expression of ITGB4 in GBM tissues (left, n = 163) and normal tissues (right, n = 207) was analyzed through GEPIA online tools. (D) The correlation between ITGB4 expression and GBM patients’ prognosis was analyzed through GEPIA online tools. *P < 0.05.
Fig 5: RhoA activity and migration of cells. (A, B) Images of the emission ratios of YFP/CFP-based RhoA biosensor in cells on normal and stiff substrate. Cells are shown on the left, the right panels show the emission ratios of the YFP/CFP-based RhoA biosensors (C) Representative images of wound-healing response for NC and ITGB4. -/- cells on normal and stiff substrate before and after CN03 treatment. (D) Track migration of NC and ITGB4-/- cells on the normal and stiff substrate before and after CN03 treatment, monitored for 24 h (n = 40). Tracks were recorded in the x/y directions. (E) Quantitation of the rate of repair at 24 h post-injury (relative to 0 h, n = 40). (F) Box-and-whisker plot of cell migration velocity (n = 40). (n = 3 independent experiments, *p < 0.05; **p < 0.01). Data are presented as mean ± SD.
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