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. 2023 Sep 25;12(10):1800. doi: 10.3390/antiox12101800

Figure 6.

Figure 6

Knockdown of DUOX2 in IPEC-J2 increased PVEC angiogenesis in vitro. (a) The mRNA level of DUOX2 in IPEC-J2 with siRNA against DUOX2. (b) The ROS level in IPEC-J2 with siRNA against DUOX2. n = 6. Images of tube formation (c,d) and cell migration (e,f) of PVEC cultured with conditioned media from the IPEC-J2 with siRNA against DUOX2. bar = 75 μm. (g) The mRNA expression levels of angiogenic factors in IPEC-J2. (h) Images of DUOX2 and MMP3 immunofluorescence staining in IPEC-J2 with siRNA against DUOX2. bar = 200 μm. (i) IPEC-J2 with siRNA against DUOX2 was treated with actinomycin D (5 μg/mL) for 0, 6, and 12 h, then RNA was isolated at indicated time points. qPCR was performed to assess the mRNA expression level of MMP3. (j) Proliferation assay of IPEC-J2 cells. IPEC-J2 with siRNA against DUOX2 were treated with or without 10 μM MMP3 inhibitor UK356618 for 24, 48, and 72 h, respectively. Images of wound healing assay (k,l) and tube formation (k,m) of PVEC cultured with conditioned media from the NC and DUOX2 knockdown cells treated with or without 10 μM UK356618. bar = 75 μm. Values are described as mean ± SEM, n = 6. The difference between the two groups was analyzed using the Student’s t-test. * indicates p < 0.05. NS, no significance. All experiments were performed in triplicate.