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. 2024 Jul 31;32(3):101311. doi: 10.1016/j.omtm.2024.101311

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© 2024 The Author(s)

This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

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Differentiation-related expression of EGFP in HaCaT cells transduced with SINγ-RVs

(A) Top: schematic of self-inactivating γ-retroviral vectors (SINγ-RVs) carrying the three different promoters that drive the expression of the bicistronic gene encoding EGFP and puromycin resistance. Bottom: graphic of the HaCaT differentiation protocol. (B) K10 (left) and endogenous TGM1 (ED TGM1) (right) relative mRNA expression in non-transduced (NT) and transduced (TGM1^full) HaCaT cells, grown in low calcium (t0) and high calcium for 1, 4, and 6 days. GAPDH mRNA was used to normalize the RT-PCR. (C) RT-qPCR quantification of the relative mRNA expression value of EGFP. GAPDH mRNA was used to normalize the RT-PCR (∗p < 0.0001). (D) Western blot analysis of EGFP expression in low-calcium (t0) and high-calcium culture conditions for 4 or 6 days of cultivation. NT HaCaT cells 6 days after calcium addition are loaded as a reference sample. t0: time zero.