Fig. 2 | Scientific Reports

Fig. 2

From: MAP4 kinase-regulated reduced CLSTN1 expression in medulloblastoma is associated with increased invasiveness

Fig. 2

Subcellular distribution of CLSTN1 in cultured MB cells. (A) Immunoblot (IB) analysis of CLSTN1 expression in six different MB cell lines. GAPDH expression was used as the loading control. (B) Surface expression analysis of CLSTN1 in six different MB cell lines by FC. The bar plot depicts mean fluorescence intensities in the six different cell lines from different experiments. (C) Subcellular fractionation analysis of CLSTN1 expression in the SHH MB cell line UW228 and the Grp3 MB cell line HD-MB03. Integrin-a5 and PRPF9 expression were used as fractionation controls for the membrane and the nuclear fractions, respectively. CE: cytoplasmic extraction; ME: membrane extraction; NE: nuclear extraction. UW228: n = 3, HD-MB03: n = 2 independent experiments. (D) IFA of endogenous CLSTN1 protein in UW228 LA-EGFP-mCherry-Nuc cells. Inverted greyscale images of the green-fluorescent signal (LA-EGFP, middle) and the red-fluorescent signal (CLSTN1, right) are shown. Scale bar is 20 µm. Magnifications a, b and c depict CLSTN1 localization in filamentous cell-cell connections and lamellipodia-like protrusions, respectively. Arrowheads indicate CLSTN1 localization in the plasma membrane, arrows accumulations of CLSTN1 in the filamentous cell-cell connections.

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