# The Tetraspanin CD151 Is Required for Met-dependent Signaling and Tumor Cell Growth

Mélanie Franco, Simona Corso, Enrico Tenaglia, Andrea Bertotti, Lorena

Capparuccia, Livio Trusolino, Paolo M Comoglio, Luca Tamagnone

► To cite this version:

Mélanie Franco, Simona Corso, Enrico Tenaglia, Andrea Bertotti, Lorena Capparuccia, et al. The Tetraspanin CD151 Is Required for Met-dependent Signaling and Tumor Cell Growth. Journal of Biological Chemistry, 2010, 285, pp.38756 - 38764. 10.1074/jbc.m110.145417. hal-05357279

HAL Id: hal-05357279

https://hal.science/hal-05357279v1

Submitted on 10 Nov 2025

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<figure><img src="image_1.png" /></figure>

Distributed under a Creative Commons Attribution 4.0 International License
SUPPLEMENTARY MATERIAL

SUPPLEMENTAL METHODS

Cell migration assays

The haptotactic migration of tumor cells on ECM substrates was analyzed using Transwell chambers (Costar). The lower side of porous filter was coated with 10µg/mL of either laminin or fibronectin. 5x10⁴ A431 cells were seeded in the upper chamber and incubated in medium with 0,1% BSA. After 18 hours, the non-migrated cells on the upper side of filters were cleared with cotton swabs, and the migrated cells were fixed with 11% glutaraldehyde and stained with crystal violet. After solubilization with 10% acetic acid, dye absorbance at 595nm was read using a microplate reader.

SUPPLEMENTAL FIGURES

Suppl. Fig. 1. CD151 is required for haptotactic migration on laminin but not on fibronectin ECM substrate. Tumor cell migration was evaluated using Transwell chamber with filters coated with either laminin (A) or fibronectin (B) in presence or absence of HGF (20ng/mL)(see Suppl. Methods for details). Data are means of two independent experiments performed in triplicate; statistical significance was assessed by comparing HGF-treated cells with respective controls in each experimental condition; *p<0.05.

Suppl. Fig. 2. Quantitative Real Time PCR analysis of CD151 levels in control and knocked-down A549 cells, or in cells rescued by over-expression of CD151.

Suppl. Fig. 3. Expression analysis of A549 carrying HGF-autocrine loop and/or shRNA targeting CD151 transcript. A549 cells were transduced with lentiviral vectors to express His-HGF and/or shRNA-CD151. CD151-silencing (A) and HGF overexpression (B) were verified by real-time Q-PCR. HGF secretion was furthermore verified by western blotting in cell-conditioned media, using anti-His-tag antibodies (C).

Suppl. Fig. 4. Quantitative Real Time PCR analysis of CD151 levels in control and A549 cells expressing shRNA-CD151 (used in most of the experiments) or an additional shRNA clone (shRNA-CD151_2) targeting a different sequence of the gene (see Methods in main manuscript).

Suppl. Fig. 5. CD151 was transfected with or without β4 integrin in COS cells expressing endogenous Met-CD151 was immunopurified from cell lysates, and the immunocomplexes were analyzed by Western blotting with specific antibodies to detect associated Met and β4 receptors. Met was co-immunoprecipitated with CD151 independently of β4 expression.
<caption>Suppl. Figure 1</caption>

<figure><img src="image_2.png" /><figcaption>B</figcaption></figure>

<figure><img src="image_3.png" /></figure>
<figure><img src="image_4.png" /><figcaption>Suppl. Figure 2</figcaption></figure>
<caption>Suppl. Figure 3</caption>

<figure><img src="image_5.png" /><figcaption>B</figcaption></figure>

<figure><img src="image_6.png" /><figcaption>C</figcaption></figure>

<figure><img src="image_7.png" /></figure>
<figure><img src="image_8.png" /><figcaption>Suppl. Figure 4</figcaption></figure>
<figure><img src="image_9.png" /><figcaption>Suppl. Figure 5</figcaption></figure>