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Cultured MECs were treated with Wnt3a-containing conditioned medium (or control), and Axin2 was found to be induced 6×even in Lrp5−/− MECs.

When slices are cocultured with reelin+ HEK cells in presence of: RAP, PP2, and LY294002, the percentage of radial glial processes expressing both markers, vimentin and BLBP, does not increase and is similar to controls (i.e., E24 MAM treated slices in plain medium or control HEK cells; Figure 4E,G,H,K).

To count synaptic contacts, 7-day-old wt, podxl and wt primary cultures incubated with conditioned medium containing PC ectodomain (30 50 µl conditioned medium in 450 µl of total medium) or control medium (rabbit Fc) were fixed and labeled with the MAP-2 and Synapsin I and II antibodies.

Cells were grown in 50% Wnt-3a conditioned medium (or control media) plus 50% normal growth media.

After 4 days in fusion medium, cells were exposed to ∼0.1 nM agrin conditioned medium or control medium for 24 h prior to cell staining and fixation.

The extent of mineralisation detected by retained alizarin red S dye was significantly lower in SaOS-2 cells treated with conditioned medium from LNCaP cells overexpressing Id-1 (grey columns, P<0.05 for each of the three clones, compared with either vector control conditioned medium or control medium).

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HUVECs were treated with Sema3C-conditioned medium or control-conditioned medium from GFP-expressing pericytes.

Muscle progenitor cells were then cultured in this heparin-depleted hESC-conditioned medium, hESC-conditioned medium, or controls (medium alone and medium conditioned by differentiated cells that lack the pro-regenerative activity).

For neurite length and branching analysis, 2 3 day-old wt, podxl and wt primary cultures incubated with conditioned medium containing PC ectodomain (30 50 µl conditioned medium in 450 µl of total medium) or with control medium (rabbit Fc) were fixed and labeled with anti-TUJ-1.

After 24 hours of exposure to conditioned medium from SC cultures (or control medium), C2C12 transcription of myogenic markers was increased only when supplemented with conditioned medium from NMD-matrix SC cultures.

SCF (10 or 100 ng/ml medium) or vehicle (controls) was added to the cells and incubated at 37°C for 5, 10 or 15 minutes.

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