Fig 2: Effects of Itgb1-knockout on the function and survival of iPSCs cultured on Matrigel. a Representative fluorescence imaging (left) and quantification (right) of PKH26-labeled iPSCs adhered to plates with agarose, collagen and Matrigel (n = 5). b Representative fluorescence imaging (left) and quantification (right) of PKH26-labeled iPSC migration across the chamber membrane in the presence of agarose, collagen and Matrigel (n = 3). c Levels of iPSCs early apoptosis in the presence of agarose, collagen and Matrigel were determined by Annexin V/FITC staining (n = 3). d iPSC proliferation in the presence of agarose, collagen and Matrigel determined by CCK-8 assay (n = 6). p < 0.05, **p < 0.01, ***p < 0.001. OD, optical density; PI, propidium iodide

Fig 3: A schematic representing the role of AZIN2-sv in angiogenesis.AZIN2-sv is an anti-angiogenesis lncRNA that acts by regulating Tln1 degradation through ubiquitination and binding to miR-214, resulting in the suppression of ITGB1 and inhibition of the Akt pathway and poor prognosis after MI. The transcription factor Bach1, a negative regulator of angiogenesis, can bind to the AZIN2-sv promoter and enhance its expression.

Fig 4: Tln1 is a downstream interactor protein of AZIN2-sv.(A) Silver-stained SDS-PAGE gel image representing proteins immunoprecipitated by AZIN2-sv and its antisense RNA in endothelial cells. The arrow indicates the region of the gel excised for western blotting. (B) Tln1 protein was detected in the specific band by western blotting. (C) RIP assays were performed using an antibody against Tln1 or negative control IgG. The purified RNA was used for qRT-PCR analysis, and enrichment of the AZIN2-sv was normalized against the input. ⁎p < .05 vs. RIP with IgG; n = 6 per group. (D) Agarose gel electrophoresis of RIP products. Arrow indicates enriched AZIN2-sv. (E) FISH and immunofluorescence for AZIN2-sv and Tln1 forty-eight hours after transfection with AZIN2-sv or control vector (bars, 40 μm). Blue indicates the nucleus, and red and green indicate the location of AZIN2-sv and Tln1, respectively. (F) ITGB1 and Tln1 protein levels detected by western blotting when AZIN2-sv was knocked down or overexpressed (GAPDH is the internal reference). (G) Quantification of protein levels in (F). ⁎p < .05 vs. vector. #p < .05 vs. shNC; n = 6 per group.

Fig 5: Effects of Itgb1-knockout on Matrigel-embedded iPSC-mediated wound healing and iPSC survival in skin wound tissues. a Representative macroscopic images of cutaneous wounds treated with Matrigel- or agarose-embedded iPSCs on days 0, 2, 4, 6, 8, 10, and 12 post-treatment (left). Quantification of wound healing expressed as the percentage of healing relative to initial wound size at day 0 (right) (n = 10). b Representative photomicrographs of CD31 IHC staining of wound tissues on days 6 and 12 post-treatment (left). CD31 signal densities were determined using ImageJ software (right) (n = 5). c Quantification of Cd34 and Cd31 mRNA levels in wound tissues (n = 10). d Representative blood perfusion images of cutaneous wounds treated with iPSCs or an equal volume of PBS on days 0, 2, 4, 6, 8, 10, and 12 post-treatment (left). Quantification of blood perfusion in wound tissues (right) (n = 10). e Representative agarose gel electrophoresis images of Tet-On and 18S DNA (left). Quantification of Tet-on expression normalized against 18S expression in wound tissues (right) (n = 10). *p < 0.05, **p < 0.01, ***p < 0.0001