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. 2020 Dec 2;148(5):1245–1259. doi: 10.1002/ijc.33383

FIGURE 2.

FIGURE 2

Modulation of SDC2 effects TGFβ signalling, migration and immunosuppressive properties of TASCs. A, RT‐qPCR analysis showing siRNA‐SDC2 transfected TASCs (siSDC2‐A and siSDC2‐B) have reduced SDC2 RNA levels compared to siControl transfected TASCs. B, siControl or siSDC2 transfected TASCs were treated with TGFβ. 2 hours after TGFβ treatment RNA was prepared and levels of SMAD7 were determined by RT‐qPCR. n = 3; ***P ≤ .0001. C, shControl (shCt) or shSDC2 transduced TASCs were treated with TGFβ. 2 hours after TGFβ treatment RNA was prepared and levels of SDC2 and TGFβ‐regulated genes were determined by RT‐qPCR. n = 4; *P ≤ .05 ***P ≤ .0001. D, The xCelligence system was used to determine the ability of shCt and shSDC2 transduced TASCs to migrate towards serum‐containing media. Levels of migration were normalised to the positive control (ie, shCt +serum). n = 4 *P ≤ .05. E, TASCs and umbilical cord MSCs (UC‐MSCs) were cocultured with CD3/CD28‐activated peripheral blood mononuclear cells (PBMCs) at a 1:50 ratio. Flow cytometry of CFSE‐labelled CD4+ T cells reveal CD3/CD28‐mediated proliferation is inhibited by TASCs. One‐way analysis of variance (ANOVA) with Tukey's multiple comparison test ***P ≤ .001. F, shCt or shSDC2 transduced TASCs were cocultured with CD3/CD28‐activated peripheral blood mononuclear cells (PBMCs) at a ratio of 1:50. Flow cytometry of CFSE‐labelled CD4+ T cells was used to measure the level of proliferation. Data were compared by one‐way ANOVA with Tukey's multiple comparison posttest. n = 4 (**P ≤ .01)