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. 2016 Dec 28;8(6):10050–10063. doi: 10.18632/oncotarget.14327

Figure 2. Effects of the overexpression of reprogramming factors after doxycycline induction.

Figure 2

a. Diagram of the single doxycycline-inducible polycistronic transgene in the Col1a1 locus, which drives the expression of the four mouse reprogramming factors (OSKM, encoding Oct4, Sox2, Klf4 and c-Myc) and b. the establishment of a long-term culture of SSCs from α-6+/c-Kit-/low testicular cells of Col1a1-4F2A-OSKM mice. c. Expression of the transgene (E2A-c-Myc, end part of the insert) in established SSCs cultures from Col1a1-4F2A-OSKM SSCs after 2 days of DOX induction (normalized to uninduced SSCs culture, n=3). *** P<0.001. d. Protocol used for cell reprogramming. e. Sorting strategy of testicular cell populations: spermatogonial populations of SP/α6+/c-Kit-/low and SP/ α6+/c-Kit+ progenitors and the spermatocyte I (4N) population. f. Bright-field microscopy showing representative pluripotent colonies obtained after induction using reprogramming factors in a SP/ α6+/c-Kit+ sorted population, and the reprogramming efficiency of the different cell populations tested.