Molecular mechanisms underlying Zeb1 regulation of alkali-induced corneal inflammation
(A) BMCs and MEFs of both Zeb1 wild type (Zeb1+/+) and Zeb1-KO (Zeb1−/+ or Zeb1−/−) were monolayer cultured and treated with or without 1 μg/mL LPS for 1 h. Total proteins were extracted from these cells for WB analyses for the indicated proteins.
(B) Chromatin immunoprecipitation (ChIP) assessments to identify Zeb1 binding to the putative promoter sequences of the indicated genes. Input, 1/10 of the initial chromatin used for immunoprecipitation; Zeb1, the anti-Zeb1 rabbit-serum-precipitated chromatin; H3, anti-pan histone 3 as a positive control; IgG, rabbit pre-serum as a negative control; Mock, without any antibodies.
(C) Zeb1 expression detected by qPCR in both BMCs and MEFs upon 25 ng/mL of TGFβ1 treatment for 16 h. Ctrl, PBS control. p-Nf-κb, phosphorylated NF-κB; p-Stat3, phosphorylated Stat3. Data are represented as mean ± standard deviation.