Fig. 2: Adipose-derived factors expand CD44v6 + cell fraction that secretes NGF and potentiates the migration capacity of ASCs.

a Cytokines secreted by CR-CSphCs (n = 4: #1, #8, #9, #21), S-ASCs (n = 6: #3, #5, #6, #8, #14, #20), V-ASCs (n = 6: #3, #5, #6, #8, #14, #18), or primary adipose tissue (AT) (n = 4). Data are the mean of 3 independent experiments. b Cell growth of CR-CSphCs treated for 5 days with IL-6 and HGF alone or in combination. The dotted red line shows the cell number at day 0. c Colony size of CR-CSphCs treated as indicated. n represents the number of colonies. Statistical significance was calculated using the two-tailed t test. d Invasion assay of CR-CSphCs pretreated with the indicated cytokines for 48 h. For b–d data show mean ± S.D. of three independent experiments using fourdifferent CR-CSphCs (CSphC #1, #8, #9, #21). e mRNA expression levels of CSC-related genes in CMS2 CR-CSphCs (CSphC #8, 9) exposed to vehicle (Medium) or IL-6 in combination with HGF for 48 h. f Immunoblot analysis of ZEB2 in CMS2 CR-CSphCs (CSphC #8) treated as indicated. Data are mean ± S.D. of three independent experiments using two different CSphCs (CSphC #8, #9). Samples were run on the same gel and images were cropped only for the purpose of this figure. Source data are provided as a Source Data file. g Kinetic growth of CR-CSphCs treated as indicated. h Number of invading CR-CSphCs at 48 h, pretreated with V-ASC CM and the indicated neutralizing antibodies for 48 h. i Flow cytometry analysis of CD44v6 positivity in CR-CSphCs treated as indicated, for 48 h. For g–i data are mean ± S.D. of six independent experiments performed with 2 different CR-CSphC lines (#8 and #9). For (b–d and g-i) statistical significance was calculated using the unpaired two-tailed t test. j, CD44v6 expression in CD44v6^- and GFP-transduced CD44v6 + cells after 3 days of exposure to V-ASC CM. One representative of 6 independent experiments is shown. k NGF, BDNF, NTF3, and NTF4 mRNA expression levels on CD44v6^- and CD44v6 + cells. Results show mean ± S.D. of three independent experiments performed with enriched cells from two different CR-CSphC lines (CSphC #8, #9). l Lollipop plot showing NGF, BDNF, NT-3, and NT-4 production by the indicated cells treated as indicated. m, Invasion assay of RFP transduced ASCs, using the indicated cells/media as chemoattractant agents. Scale bars, 100 µm. n Number of invading ASCs in presence of the indicated cells/media as chemoattractant agents. For l–n data are mean ± SD of three independent experiments using CR-CSphCs from different patients (CSphC #1, #8, #9, #21).