Skip to main content
. 2023 May 13;6:516. doi: 10.1038/s42003-023-04905-z

Fig. 5. JunB regulates IL-33-induced IL-8 promoter activity.

Fig. 5

a IL-8 promoter encompassing from −342 to +59 nt (401 kb) was cloned into pGL3-basic vector and sequenced. b HRMVECs were transfected with pGL3-basic vector or pGL3-IL-8 promoter (401 kb), growth-arrested, and treated with or without IL-33 (20 ng/mL) for 6 h, and the luciferase activities were measured. c Quiescent HRMVECs were treated with or without IL-33 (20 ng/mL) for various time periods and analysed by Western blotting for indicated proteins and normalized to β-tubulin. d HRMVECs were transfected with control siRNA (siControl) or VCAM-1 siRNA (siVCAM-1), quiesced, treated with or without IL-33 (20 ng/mL) for 6 h, cell extracts were prepared and analysed by Western blotting for JunB levels, and the blot was reprobed for VCAM-1 and β-tubulin to show the effects of siRNA on its target and off target molecules. e HRMVECs were transfected with pGL3-basic vector or pGL3-IL-8 promoter (1.215 kb) in combination with control siRNA (siControl) or JunB siRNA (siJunB), growth-arrested, and treated with and without IL-33 (20 ng/mL) for 6 h, and the luciferase activities were measured. f HRMVECs were transfected with control siRNA (siControl) or JunB siRNA (siJunB), quiesced, treated with or without IL-33 (20 ng/mL) for 8 h, cell extracts were prepared and analysed by Western blotting for the indicated proteins. g C57BL/6 mice pups were exposed to 75% oxygen and at various time periods of relative hypoxia, eyes were enucleated, retinas isolated, tissue extracts made and analysed by Western blotting for JunB levels using their specific antibodies and normalized to β-tubulin. h At P15, the retinal tissue extracts from IL-33+/+ and IL-33−/− mice pups subjected to normoxia and hypoxia were analyzed for indicated proteins by Western blotting. n = 6 (b, e) or n = 3 (c, g) biologically independent samples per group, expressed as Mean ± SD. *P < 0.05 vs normoxia or control or siControl, **p < 0.05 vs siControl + IL-33.