GADD45B Antibody - #DF2375

Fig. 2. GADD45B aggravated inflammation and cellular senescence in CSE-induced HBE cells. a, b RNA-sequencing of HBE cells with lentivirus infections was performed (n = 3 per group). Volcano plot showed the DEGs in HBE cells infected with Lv-GADD45B compared with Lv-GFP. The results of biological process enrichment analysis of DEGs. c Western blot analysis of Flag in HBE cells infected with Lv-GADD45B (n = 4). d ELISA analysis of IL-6, IL-8 and IL-1β levels in CSE-induced HBE cells with or without Lv-GADD45B infection (n = 7). e–g Western blot analysis of P16 and P21 expressions in HBE cells infected with Lv-GADD45B under CSE treatment (n = 3). h, i Representative images of SA-β-gal staining and percentages of SA-β-gal positive cells for CSE-treated HBE cells overexpressing GADD45B (n = 5, arrowheads indicated the positively stained cells). Scale bar = 50 μm, magnification = x 200. j Western blot analysis for GADD45B expression in HBE cells with siRNA transfection (n = 4). k ELISA analysis of IL-6, IL-8 and IL-1β levels in CSE-induced HBE cells with or without si-GADD45B transfection (n = 4). l–n Western blot analysis of P16 and P21 expressions in HBE cells transfected with si-GADD45B under CSE treatment (n = 3). o, p Representative images of SA-β-gal staining and percentages of SA-β-gal positive cells for CSE-treated HBE cells with GADD45B knockdown (n = 5, arrowheads indicated the positively stained cells). Scale bar = 50 μm, magnification = x 200. Data were expressed as mean ± SD. P-values were calculated using one-way ANOVA followed by Newman–Keuls test or Student’s unpaired t-test. *P < 0.05, **P < 0.01, and ***P < 0.001 represent significant differences. GADD45B, growth arrest and DNA damage-inducible 45 beta; CSE, cigarette smoke extract; HBE, human bronchial epithelial; Lv-GADD45B, lentivirus expressing GADD45B; Lv-GFP, lentivirus with empty vector carrying green fluorescent protein; DEGs, differentially expressed genes; SA-β-gal, senescence-associated β-galactosidase; siRNA, small interfering RNA; NC, negative control. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Figure 3 GADD45B knockdown inhibited the migration of EOC cells. (A, B) Western blot and qRT-PCR detected the expression of GADD45B in the EOC cell lines. (C, D) qRT-PCR and Western blot analysis of the silencing efficiency of GADD45B. (E, G) Wound-healing assays detected the migration ability difference of ES2 and SKOV3 cells between the NC group and the GADD45B siRNA-downregulated groups. (F, H) Histograms show the wound-healing areas of the scratch-wound assays. (I) Transwell assays detected the migration ability of ES2 and SKOV3 cells in the NC group and the GADD45B siRNA-downregulated groups. (J, K) Histograms show the number of migrating cells. Experiments were performed at least in triplicate, and the results are shown as the mean ± s.d. Student’s t-test was used to analyse the data (*p < 0.05; **p < 0.01; ***p < 0.001). Scale bar, 100 μm.

Figure 7 GADD45B is overexpressed in omental metastatic tissues of EOC over matched primary tumour tissues. (A) Representative images of the immunohistochemical analysis of GADD45B in one of the paired tumour tissues. Scale bar, 20 μm. (B) Statistical result of the mean density of GADD45B in 11 pairs of matched EOC tissue samples. Paired Student’s t-test was used to analyse the data (**p < 0.01).

Fig. 5 Shikonin increased tumor suppressor genes PPP3CC and GADD45B in MAPK signaling. A-B. GADD45B and PPP3CC are activated after shikonin treatment in both A549 and H1299 cells by qPCR detection. C-D. The MAPK proteins, GADD45B and PPP3CC are activated after shikonin treatment in both A549 and H1299 cells by western blotting. The blots are cut prior to hybridisation with antibodies during blotting and the cropped plots are used in the figure to improve the clarity of presentation.