Fig. 2: Lineage- and cell type-independent role for AJSZ as barriers to reprogramming. | Nature Communications

Fig. 2: Lineage- and cell type-independent role for AJSZ as barriers to reprogramming.

From: Conserved transcription factors promote cell fate stability and restrict reprogramming potential in differentiated cells

Fig. 2

a Expression of AJSZ in HDFs. Immunostaining for AJSZ is shown in red and fibroblast markers Vimentin (VIM) or Transgelin (TAGLN) are shown in green. Nuclei are stained with DAPI. n = 4 per condition. b Quantification of the % of cardiac marker ACTN2-expressing cells normalized to MGT + siControl condition after 30 days of cardiac reprogramming in HDFs. n = 5 and n = 9 for siControl and siAJSZ treated, respectively. c Representative images of HDFs treated with MGT + siControl or MGT + siAJSZ and immunostained for cardiac (ACTN2, green) and fibroblast (TAGLN, red) markers. Nuclei are stained with DAPI (blue). White arrows indicate iCMs. d Quantification of the % of MAP2 (green) and TUJ1 (red)-double-positive cells, 3 days after overexpression of Ascl1, Brn2, and Mytl1 (ABM) in HDFs. n = 7 and n = 8 for siControl and siAJSZ treated, respectively. e Representative images of HDFs treated with ABM + siControl or ABM + siAJSZ and immunostained for MAP2 (green) and TUJ1 (red). Nuclei are stained with DAPI (blue, top left insets). White arrows indicate TUJ1 + MAP2 + double-positive cells = neurons. f Quantification of the % of NANOG-positive cells, 7 days after overexpression of OCT4, KLF4, SOX2, and cMYC (OKSM) in HDFs. n = 10 per condition. g Representative images of HDFs treated with OKSM + siControl or OKSM + siAJSZ and immunostained for pluripotent marker NANOG (red). Nuclei are stained with DAPI (blue, top left insets). White arrows indicate NANOG-positive cells= iPSCs. h Quantification of the % of ACTN2 + 20 days after MGT overexpression. i Representative images of HAECs treated with MGT + siControl or MGT + siAJSZ and immunostained for the endothelial marker (PECAM1, red) and the cardiac marker (ACTN2, green). n = 4 per condition. Nuclei are stained with DAPI (blue). White arrows indicate iCMs. Scale bars: 50 µm. Student’s t-test: *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001. Groups were compared using two-tailed unpaired analysis. Data in figure are presented as mean values ± standard deviation.

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