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. Author manuscript; available in PMC: 2018 Mar 7.
Published in final edited form as: Pigment Cell Melanoma Res. 2017 Mar 7;30(2):203–218. doi: 10.1111/pcmr.12564

Figure 6. Exosomes from B16F0 but not Cloudman S91 or Melan-A cell lines contained PTPN11 protein and were enriched for PTPN11 mRNA.

Figure 6

(a) Presence of PTPN11 and the exosome markers CD63 and CD9 were assayed in whole cell lysates (WCL) and exosome (EXO) samples from B16F0 cells. (b) The presence of PTPN11 and the exosome markers CD63 and HSP70 were assayed in whole cell lysates and exosome samples from Melan-A, Cloudman S91, and B16F0 cell lines. In Panels (a) and (b), β-actin was included as a negative control for the exosome samples and 20 µg of total protein was loaded in each lane. (c and d) RT-PCR was used to quantify within equally loaded cDNA samples the abundance of transcripts associated with Gapdh, Ptpn11, Rnd2, Eif4abp2, and Wsb2 in cells (c) and in exosomes (d). In (c), relative gene expression is reported using ΔΔCt, where the numerator is the ΔCt for the indicated gene relative to Gapdh in the sample and the numerator is the ΔCt for the average Cts of the indicated gene relative to Gapdh in the B16F0 sample. In (d), relative gene expression is reported using ΔCt for the indicated gene in the exosome sample relative to the cell sample. Statistical differences were assessed using Students t-test, where ** and *** correspond to p < 0.05 and p < 0.001, respectively, for a sample compared to the corresponding B16F0 sample.