Fig 1: Promotion of GBM migration by migrasomes via PAK4 and LAMA4.a NC, Kd-LAMA4, and Kd-PAK4 cells were stained with WGA and the indicated antibodies and visualized using confocal microscopy. Scale bar, 10 μm. High magnification images of the migrasomes are shown in the white boxes. Scale bar, 2 μm. b Statistics of the amount of PAK4/ LAMA4 released into the supernatant by cells in the NC and Kd-TSPAN4 groups. ****P < 0.0001. c The staining of cells migrating to the bottom of the chambers in the NC and Kd-PAK4 groups after adding different migrasomes was observed using crystal violet. Scale bar, 50 μm. d The staining of cells migrating to the bottom of the chambers in the NC and Kd-LAMA4 groups after adding different migrasomes was observed using crystal violet. Scale bar, 50 μm.
Fig 2: ECM-related factors contained in GBM migrasomes.a Volcano plot of the quantification of TMT-labelled proteins through mass spectrometry. Red dots indicate a migrasome with a cell abundance ratio of >2 (P < 0.05), while blue dots indicate a migrasome with a cell abundance ratio of <0.5 (P < 0.05). The experiment was performed independently three times (n = 3). PAK4 and LAMA4 are labeled separately. A two-tailed, two-sample, unequal variance t-test was used in Excel to calculate the P values. b, d The abundance of the indicated proteins in 5a was analyzed. P values were calculated using a one-tailed, two-sample unequal variance t-test. The P values were as follows: histone (P = 0.000215), integrin (P = 0.000379), tetraspanin (P = 0.000556), cytokines (P = 0.001711), PAK4 (P = 0.001885), LAMA4 (P = 0.008333), and ELP6 (P = 0.000181). c For the gene ontology (GO) enrichment analysis at the biological process (BP) level of the red-dot proteins in 5a, the pathways associated with migration are highlighted in red. e Western blot was used to analyze cell body and migrasome proteins with anti-LAMA4 and anti-PAK4 antibodies. Two batches of cell body and migrasome proteins were extracted. CB: cell body. Mig: migrasome. f LN229 cells were visualized using confocal microscopy after staining with WGA, anti-LAMA4, and anti-PAK4 antibodies. Scale bar, 10 μm. High magnification images of the migrasomes are shown in the white boxes. Scale bar, 2 μm. For the co-localization analysis of the boxed region, the arrows indicate co-localization. g Staining of cells migrating/invading to the bottom of Transwell chambers using crystal violet. Cells were treated with PBS or recombinant proteins PAK4 and LAMA4. ***P < 0.001. ****P < 0.0001.
Supplier Page from Fine Biotech Co., Ltd. for Recombinant Human PAK4