1 lg anti epcam antibody proteintech  (Proteintech)

Journal: Redox Biology

Article Title: Ionizing radiation promotes lung injury by inducing ferroptosis-driven senescence in epithelial cells via NCOA4-mediated ferritinophagy

doi: 10.1016/j.redox.2026.104091

Figure Lengend Snippet: RILI is associated with lung epithelial cells ferroptosis and senescence. A. HE and Masson staining were used to evaluate RILI (Scale bar, 100 μm). B. IHC revealed decreased GPX4 expression in lung tissue at 4 weeks after IR exposure (scale bars, 100 μm and 20 μm). C. The levels of GPX4 expression at different times after IR exposure were measured by Western blot analysis. D-G. Bar graph showing the levels of tissue and serum iron content (D, E), MDA (F), GSH/GSSG ratio (G) in lung tissues at different times after IR exposure. n = 6 for each group. H . Immunofluorescence detection of epithelial cell senescence in lung tissue at different times after IR exposure using anti-EpCAM (green) and anti-P16 (red) antibodies (Scale bar, 100 μm). I. The mRNA expression of senescence markers (P16, P21) and SASP factors (MMP9, CCL2, CCL7, and CXCL1) was analyzed by RT-qPCR in the lung tissue at different times after IR exposure. n = 3 for each group. J. The protein expression of P16 and P21 was detected by Western blot in the lung tissue at different times after IR exposure. K. Representative images (left) and relative fluorescence intensity quantitative analysis (right) of Fe 2+ levels in BEAS-2B cells stained with FerroOrange after exposure to 0, 2, 5, and 10 Gy IR for 24 h (scale bar, 100 μm). n = 3 for each group. L. The intracellular level of ROS was detected by flow cytometry, and the mean fluorescence intensity was subsequently analyzed. n = 3 for each group. M. The levels of GPX4 expression were measured 24 h post-IR using Western blot analysis at different radiation doses. N–P. Bar graph showing the levels of iron content (N), MDA (O) and GSH/GSSG ratio (P) in the BEAS-2B cells 24 h post exposure to 0, 2, 5, and 10 Gy IR. n = 3 for each group. Q. Representative images of SA-β-gal activity in BEAS-2B cells 48 h post exposure to 0, 2, 5, and 10 Gy IR (Scale bar, 100 μm). R. RT-qPCR was used to analyze the mRNA expression of senescence markers (P16, P21) and SASP factors (MMP9, CCL2, CCL7, and CXCL1) in BEAS-2B cells at different times after exposure to 10 Gy of IR. n = 3 for each group. S. The levels of P16 and P21 expression were measured by Western blot at different times after 10 Gy IR. ns, p ≥ 0.05; ∗, p < 0.05; ∗∗, p < 0.01; ∗∗∗, p < 0.001.

Article Snippet: The next day, species-specific fluorescent secondary antibodies (1:200, Elabscience, China) were applied and incubated at 37 °C for 1 h. The detection of epithelial cell senescence in lung tissue is performed using anti-EpCAM (1:500; Proteintech, 21050-1-AP, China) along with anti-CDKN2A (P16, 1:200; Santa Cruz Biotechnology, sc-1661, USA).

Techniques: Staining, Expressing, Western Blot, Immunofluorescence, Quantitative RT-PCR, Fluorescence, Flow Cytometry, Activity Assay

Journal: Redox Biology

Article Title: Ionizing radiation promotes lung injury by inducing ferroptosis-driven senescence in epithelial cells via NCOA4-mediated ferritinophagy

doi: 10.1016/j.redox.2026.104091

Figure Lengend Snippet: Inhibition of ferroptosis alleviated RILI by mitigate lung epithelial cells senescence. A. HE and Masson staining reveal that ferroptosis inhibitor Fer-1 (2 mg/kg) improves RILI in mice (scale bar, 100 μm). B–C. IHC (B) and Western blot (C) revealed that the ferroptosis inhibitor Fer-1 restored GPX4 expression in lung tissue at 4 weeks after IR exposure (scale bars, 100 μm). D-G. Bar graph showing the levels of tissue and serum iron content (D, E), MDA (F), GSH/GSSG ratio (G) in lung tissues at 4 weeks after Fer-1 treatment and IR exposure. n = 6 for each group. H . Immunofluorescence detection of epithelial cell senescence in lung tissue using anti-EpCAM (green) and anti-P16 (red) antibodies after Fer-1 treatment and IR exposure (Scale bar, 100 μm). I. The mRNA expression of senescence markers (P16, P21) and SASP factors (MMP9, CCL2, CCL7, and CXCL1) was analyzed by RT-qPCR in the lung tissue at 4 weeks after Fer-1 treatment and IR exposure. n = 3 for each group. J. The protein expression of P16 and P21 was detected by Western blot in the lung tissue at 4 weeks after Fer-1 treatment and IR exposure. K. Representative images (left) and relative fluorescence intensity quantitative analysis (right) of Fe 2+ levels in BEAS-2B cells stained with FerroOrange after exposure to Fer-1 (10 μM) pretreatment and 10 Gy IR for 24 h (scale bar, 100 μm). n = 3 for each group. L. The intracellular level of ROS was detected by flow cytometry after Fer-1 pretreatment and 10 Gy IR, and the mean fluorescence intensity was subsequently analyzed. n = 3 for each group. M. Lipid peroxidation was detected by C11-BODIPY581/591. Reductive (red) and Oxidative (green) respectively (scale bar, 100 μm). N. GPX4 expression levels were measured using Western blotting at 24 h after Fer-1 pretreatment and 10 Gy IR. O-Q. Bar graph showing the levels of iron content (O), MDA (P), and GSH/GSSG ratio (Q) in the BEAS-2B cells 24 h after Fer-1 pretreatment and 10 Gy IR. n = 3 for each group. R. Representative images of SA-β-gal activity in BEAS-2B cells 48 h after Fer-1 pretreatment and 10 Gy IR (Scale bar, 100 μm). S. RT-qPCR was used to analyze the mRNA expression of senescence markers (P16, P21) and SASP factors (MMP9, CCL2, CCL7, and CXCL1) in BEAS-2B cells 24 h after Fer-1 pretreatment and 10 Gy IR. n = 3 for each group. T. The levels of P16 and P21 expression were measured by Western blot at 48 h after Fer-1 pretreatment and 10 Gy IR. ns, p ≥ 0.05; ∗, p < 0.05; ∗∗, p < 0.01; ∗∗∗, p < 0.001.

Article Snippet: The next day, species-specific fluorescent secondary antibodies (1:200, Elabscience, China) were applied and incubated at 37 °C for 1 h. The detection of epithelial cell senescence in lung tissue is performed using anti-EpCAM (1:500; Proteintech, 21050-1-AP, China) along with anti-CDKN2A (P16, 1:200; Santa Cruz Biotechnology, sc-1661, USA).

Techniques: Inhibition, Staining, Western Blot, Expressing, Immunofluorescence, Quantitative RT-PCR, Fluorescence, Flow Cytometry, Activity Assay

Journal: Redox Biology

Article Title: Ionizing radiation promotes lung injury by inducing ferroptosis-driven senescence in epithelial cells via NCOA4-mediated ferritinophagy

doi: 10.1016/j.redox.2026.104091

Figure Lengend Snippet: Compound 9a mitigates RILI by suppressing ferritinophagy, ferroptosis, mitochondrial dysfunction and lung epithelial cells senescence. A. Schematic of timeline for IR (3 × 8 Gy) and 9a treatment in C57BL/6J mice. The control and 9a groups received equivalent volumes of PBS or 9a (10 mg/kg, intraperitoneal injection) on days 2, 4, 6, and 8 following irradiation. B. Representative images of HE and Masson-stained lung tissue from PBS or 9a-treated mice at 4 weeks after IR (Scale bar, 100 μm). C. Representative IHC images of lung tissue showing GPX4 expression (Scale bar, 100 μm). D. Immunofluorescence detection of ferritinophagy in lung tissue using anti-FTH (green) and anti-LAMP2 (red) antibodies after IR exposure and 9a treatment (Scale bar, 100 μm). E. The levels of NCOA4, FTH and GPX4 expression in lung tissue were measured by Western blot from PBS or 9a-treated mice at 4 weeks after IR. F–I. Bar graph showing the levels of tissue and serum iron content (F, G), MDA (H), GSH/GSSG ratio (I) in lung tissues from PBS or 9a-treated mice at 4 weeks after IR. n = 6 for each group. J. Representative TEM images of lung epithelial cell in lung tissue showing the morphology and structure of mitochondria from PBS or 9a-treated mice at 4 weeks after IR. (magnification: 8000 × , scale bar, 1 μm; magnification: 20000 × , scale bar, 500 nm). K. Immunofluorescence detection of epithelial cell senescence in lung tissue using anti-EpCAM (green) and anti-P16 (red) antibodies after IR exposure and 9a treatment (Scale bar, 100 μm). L. The mRNA expression of senescence markers (P16, P21) and SASP factors (MMP9, CCL2, CCL7, and CXCL1) was analyzed by RT-qPCR in the lung tissue at 4 weeks after IR exposure and 9a treatment. n = 3 for each group. M. The protein expression of P16 and P21 was detected by Western blot in the lung tissue at 4 weeks after IR exposure and 9a treatment. ns, p ≥ 0.05; ∗∗, p < 0.01; ∗∗∗, p < 0.001.

Article Snippet: The next day, species-specific fluorescent secondary antibodies (1:200, Elabscience, China) were applied and incubated at 37 °C for 1 h. The detection of epithelial cell senescence in lung tissue is performed using anti-EpCAM (1:500; Proteintech, 21050-1-AP, China) along with anti-CDKN2A (P16, 1:200; Santa Cruz Biotechnology, sc-1661, USA).

Techniques: Control, Injection, Irradiation, Staining, Expressing, Immunofluorescence, Western Blot, Quantitative RT-PCR