mle 12 mouse alveolar epithelial cell line  (ATCC)

Journal: Cancer Research

Article Title: ACSL5 Mediates Adaptation to the Palmitic Acid–Enriched Pulmonary Microenvironment to Enhance Metastatic Breast Cancer Cell Survival and Lung Metastasis

doi: 10.1158/0008-5472.CAN-25-0866

Figure Lengend Snippet: Exo-derived USP47 mediates fatty acid synthesis in AT2. A, MLE-12 cells were cocultured with exosomes derived from the control (shNC) and USP47-knockdown (shUSP47) MDA-MB-231/LM3. YAP, FASN and ACLY proteins, as well as the PA levels ( n = 5), in MLE-12 cells were detected. B, MLE-12 cells were cocultured with exosomes derived from the control (vector) and USP47-overexpression (oeUSP47) MDA-MB-231/PRI. YAP, FASN, and ACLY protein levels, as well as the PA levels ( n = 5), in MLE-12 cells were detected. C, Left, Western blotting was performed to detect YAP, FASN, and ACLY protein levels in the control (shNC) or YAP-knockdown (shYAP) MLE-12. Right, relative PA levels ( n = 5) in MLE-12 cells were detected. D, MLE-12 cells were cocultured with the control exosomes (vector) or USP47-overexpression exosomes (oeUSP47) derived from MDA-MB-231/PRI, or YAP-knockdown MLE-12 cells and its controls were cocultured with USP47-overexpression exosomes (oeUSP47) derived from MDA-MB-231/PRI. FASN and ACLY protein levels, as well as the PA levels ( n = 5), in MLE-12 cells were detected. E, MLE-12 cells were cocultured with the control exosomes (shNC) or USP47-knockdown exosomes (shUSP47) derived from MDA-MB-231/LM3, or YAP-overexpression (oeYAP) and its control MLE-12 cells (Vector) were cocultured with USP47-knockdown exosomes (shUSP47) derived from MDA-MB-231/LM3, respectively. FASN and ACLY protein levels, as well as the PA levels ( n = 5), in MLE-12 cells were detected. F and G, Relative PA levels in MLE-12 in indicated group ( n = 5). H, Relative PA levels ( n = 5) in the lung interstitial fluid of BALB/c nude mice injected with the control (shNC) or USP47-knockdown (shUSP47) MDA-MB-231/LM3 cells. I, Representative BLI and metastatic lungs of BALB/c nude mice injected with the control (shNC) or USP47-knockdown (shUSP47) MDA-MB-231/LM3 cells. J, Quantification of BLI and lung metastatic nodules ( n = 7). All data represent the mean ± SD. One-way ANOVA ( A , C–G , H , and J ) and Student t test ( B ) were utilized. *, P < 0.05; **, P < 0.01; ***, P < 0.001.

Article Snippet: Triple-negative breast cancer cell lines 4T1, MDA-MB-231, and HCC1806 and mouse alveolar epithelial cell line MLE-12 were purchased from the ATCC.

Techniques: Derivative Assay, Control, Knockdown, Plasmid Preparation, Over Expression, Western Blot, Injection

Journal: International journal of radiation biology

Article Title: Blocking MSI2 alleviated radiation-induced pulmonary fibrosis through inhibiting epithelial-mesenchymal transition.

doi: 10.1080/09553002.2025.2451617

Figure Lengend Snippet: Figure 2. The effect of radiation on the expression changes of EMT markers and MSI2 molecule in alveolar epithelial cells at different time points is depicted. (A) MLE-12 cells were exposed to an 8 Gy irradiation dose, and the expression changes of EMT-related molecules and the MSI2 molecule at various time points were detected by WB. (B-E) Statistical analysis of the relative expression grayscale values of E-cadherin, Vimentin, N-cadherin, and MSI2 proteins. (F) RLE-6TN cells were subjected to an 8 Gy irradiation dose, and the expression changes of EMT-related molecules and the MSI2 molecule at various time points were examined by WB. (G-J) Statistical analysis of the relative expression grayscale values of E-cadherin, Vimentin, N-cadherin, and MSI2 proteins. Data are presented as mean ± SEM, *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001, compared to the control group.

Article Snippet: Cells of the mouse alveolar type II epithelial cell line (MLE-12) and the rat alveolar type II epithelial cell line (RLE-6TN) were procured from the American Type Culture Collection (ATCC).

Techniques: Expressing, Irradiation, Control

Journal: International journal of radiation biology

Article Title: Blocking MSI2 alleviated radiation-induced pulmonary fibrosis through inhibiting epithelial-mesenchymal transition.

doi: 10.1080/09553002.2025.2451617

Figure Lengend Snippet: Figure 3. The inhibitory effect of MSI2 knockdown on radiation-induced EMT changes in alveolar epithelial cells is presented. (A) Morphological changes of MLE-12 cells in different treatment groups post-irradiation. (B) Morphological changes of RLE-6TN cells in different treatment groups post-irradiation. (C, E-H) Expression of EMT-related molecules and the MSI2 molecule in MLE-12 cells under different treatment conditions post-irradiation, detected by WB, along with statistical analysis of the grayscale values of the corresponding protein expression. (D, I-L) Expression of EMT-related molecules and the MSI2 molecule in RLE-6TN cells under different treatment conditions post-irradiation, detected by WB, along with statistical analysis of the grayscale values of the corresponding protein expression. Data are presented as mean ± SEM, ns P > 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001.

Article Snippet: Cells of the mouse alveolar type II epithelial cell line (MLE-12) and the rat alveolar type II epithelial cell line (RLE-6TN) were procured from the American Type Culture Collection (ATCC).

Techniques: Knockdown, Irradiation, Expressing

Journal: International journal of radiation biology

Article Title: Blocking MSI2 alleviated radiation-induced pulmonary fibrosis through inhibiting epithelial-mesenchymal transition.

doi: 10.1080/09553002.2025.2451617

Figure Lengend Snippet: Figure 4. The mechanistic role of MSI2 in radiation-induced epithelial-mesenchymal transition (EMT) in alveolar epithelial cells is presented. (A) a volcano plot of differential gene expression highlights gene variability in response to radiation. (B) a hierarchical clustering heat map depicting the expression patterns of differ entially expressed genes post-radiation. (C) a bubble chart showcasing the KEGG enrichment analysis of upregulated genes associated with radiation-induced EMT. (D) Bubble chart illustrating the KEGG enrichment analysis of downregulated genes associated with radiation-induced EMT. (E) Using WB to examine the temporal expression changes of EMT-related signaling pathway markers in MLE-12 cells at various time points following an 8 Gy irradiation dose. (F) Using WB to assess the temporal expression changes of EMT-related signaling pathway markers in RLE-6TN cells at various time points after an 8 Gy irradiation dose.

Article Snippet: Cells of the mouse alveolar type II epithelial cell line (MLE-12) and the rat alveolar type II epithelial cell line (RLE-6TN) were procured from the American Type Culture Collection (ATCC).

Techniques: Gene Expression, Expressing, Irradiation

Journal: International journal of radiation biology

Article Title: Blocking MSI2 alleviated radiation-induced pulmonary fibrosis through inhibiting epithelial-mesenchymal transition.

doi: 10.1080/09553002.2025.2451617

Figure Lengend Snippet: Figure 5. The significance of ZEB1 in the MSI2-downstream radiation-induced EMT of alveolar epithelial cells is demonstrated. (A, B) Alveolar epithelial cells were exposed to an 8 Gy irradiation dose, and the protein expression of various transcription factors was assessed at different time points using WB. (C, D) Alveolar epithelial cells were exposed to an 8 Gy irradiation dose, and the mRNA expression levels of ZEB1 at various time points post-irradiation were assessed using PCR. (E) Using WB to detect the impact of ZEB1 overexpression and MSI2 knockdown on the expression of EMT markers in radiation-induced MLE-12 cells. (F) Morphological changes of MLE-12 cells in different groups were observed under a phase-contrast microscope. Data are presented as mean ± SEM, ns P > 0.05, *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001.

Article Snippet: Cells of the mouse alveolar type II epithelial cell line (MLE-12) and the rat alveolar type II epithelial cell line (RLE-6TN) were procured from the American Type Culture Collection (ATCC).

Techniques: Irradiation, Expressing, Over Expression, Knockdown, Microscopy

Journal: Cell Communication and Signaling : CCS

Article Title: Cadmium exposure triggers alveolar epithelial cell pyroptosis by inducing mitochondrial oxidative stress and activating the cGAS-STING pathway

doi: 10.1186/s12964-024-01946-7

Figure Lengend Snippet: Pyroptosis is the primary mode of AEC death induced by cadmium. MLE12 cells were treated with different concentrations of CdCl 2 for 24 h. Cell morphology was photographed through a microscope (A, bar = 100 μm). Cell viability was detected by CCK8 assay (B, n = 5). The LDH activity of the cell culture supernatant was assayed (C, n = 3). The Pan-caspase inhibitor (ZVAD-FMK) but not the necroptosis or ferroptosis inhibitor restored cell viability within 24 h of CdCl 2 incubation (D, n = 5). Cell morphology was photographed through a microscope (E, bar = 100 μm). The expressions of SPC, NLRP3, pro-Caspase1, Caspase1 p20, IL-1β p17, and GSDMD-N protein in MLE12 cells were detected using western blot (F-G, n = 3). The fluorescence intensity of GSDMD was detected by immunofluorescence (H, bar = 5 μm). Data are expressed as the mean ± SD. Differences among multiple groups were performed using ANOVA. Tukey’s test was used as a post hoc test for pairwise comparisons. * P < 0.05, ** P < 0.01, and *** P < 0.001

Article Snippet: Mouse alveolar epithelial cell lines MLE12 were purchased from ATCC (USA, CRL-2110).

Techniques: Microscopy, CCK-8 Assay, Activity Assay, Cell Culture, Incubation, Western Blot, Fluorescence, Immunofluorescence

Journal: Cell Communication and Signaling : CCS

Article Title: Cadmium exposure triggers alveolar epithelial cell pyroptosis by inducing mitochondrial oxidative stress and activating the cGAS-STING pathway

doi: 10.1186/s12964-024-01946-7

Figure Lengend Snippet: Cadmium activates the cGAS-STING pathway in AECs. Differentially expressed genes in the MLE12 cells of Control and CdCl 2 groups ( n = 3). PCA analysis (A). Volcano plot showing the changes of MLE12 genes (fold change ≥ 2) (B), GO enrichment analysis for differentially expressed genes (C). KEGG enrichment analysis for differentially expressed genes (D). The gene expression levels of the cGAS-STING pathway (E, n = 3). The expressions of cGAS and STING protein in the lung were detected using western blot (F-G, n = 3). The expressions of cGAS and STING protein and the phosphorylation level of TBK1 in MLE12 cells were detected using western blot (H-I, n = 3). The fluorescence intensity of STING was detected by immunofluorescence (J, bar = 5 μm). Data are expressed as the mean ± SD. Differences among multiple groups were performed using ANOVA. Tukey’s test was used as a post hoc test for pairwise comparisons. * P < 0.05 and *** P < 0.001

Article Snippet: Mouse alveolar epithelial cell lines MLE12 were purchased from ATCC (USA, CRL-2110).

Techniques: Control, Gene Expression, Western Blot, Phospho-proteomics, Fluorescence, Immunofluorescence

Journal: Cell Communication and Signaling : CCS

Article Title: Cadmium exposure triggers alveolar epithelial cell pyroptosis by inducing mitochondrial oxidative stress and activating the cGAS-STING pathway

doi: 10.1186/s12964-024-01946-7

Figure Lengend Snippet: Cadmium exposure induces pyroptosis in AECs by activating the cGAS-STING pathway. MLE12 cells were treated with CdCl 2 for 24 h after RU.521 intervention for 30 min. The expression of STING protein and the phosphorylation level of TBK1 and NF-κB p65 in MLE12 cells were detected using Western blot (A-B, n = 3). The expression of pro-Il-1β mRNA in MLE12 cells was detected using Real-time PCR (C, n = 3). The expressions of NLRP3, pro-Caspase1, Caspase1 p20, pro-IL-1β, IL-1β p17, and GSDMD protein in MLE12 cells were detected using western blot (D-E, n = 3). The fluorescence intensity of GSDMD was detected by immunofluorescence (F, bar = 5 μm). Data are expressed as the mean ± SD. Differences among multiple groups were performed using ANOVA. Tukey’s test was used as a post hoc test for pairwise comparisons. * P < 0.05, ** P < 0.01, and *** P < 0.001

Article Snippet: Mouse alveolar epithelial cell lines MLE12 were purchased from ATCC (USA, CRL-2110).

Techniques: Expressing, Phospho-proteomics, Western Blot, Real-time Polymerase Chain Reaction, Fluorescence, Immunofluorescence

Journal: Cell Communication and Signaling : CCS

Article Title: Cadmium exposure triggers alveolar epithelial cell pyroptosis by inducing mitochondrial oxidative stress and activating the cGAS-STING pathway

doi: 10.1186/s12964-024-01946-7

Figure Lengend Snippet: Mitochondrial oxidative stress induced by cadmium exposure activates the cGAS-STING pathway to induce pyroptosis in AECs. MLE12 cells were treated with CdCl 2 for 24 h after mitoQ intervention for 30 min. Mitochondrial membrane potential was measured by JC-1 staining (A, bar = 50 μm). ROS and mitoROS were detected by fluorescent probes (B, bar = 100 μm). Cytosolic mtDNA ND1 and Dloop2 levels in MLE12 (C, n = 3). The expressions of pro-Il-1β and Gsdmd mRNA in MLE12 cells were detected using Real-time PCR (D, n = 3). The expressions of NLRP3, Caspase-1 p20, IL-1β p17, GSDMD, and STING protein and the phosphorylation level of TBK1 and NF-κB p65 in MLE12 cells were detected using western blot (E–F, n = 3). The fluorescence intensity of STING was detected by immunofluorescence (G, bar = 5 μm). The fluorescence intensity of GSDMD was detected by immunofluorescence (H, bar = 5 μm). Data are expressed as the mean ± SD. Differences among multiple groups were performed using ANOVA. Tukey’s test was used as a post hoc test for pairwise comparisons. * P < 0.05, ** P < 0.01, and *** P < 0.001

Article Snippet: Mouse alveolar epithelial cell lines MLE12 were purchased from ATCC (USA, CRL-2110).

Techniques: Membrane, Staining, Real-time Polymerase Chain Reaction, Phospho-proteomics, Western Blot, Fluorescence, Immunofluorescence

Journal: Cell Communication and Signaling : CCS

Article Title: Cadmium exposure triggers alveolar epithelial cell pyroptosis by inducing mitochondrial oxidative stress and activating the cGAS-STING pathway

doi: 10.1186/s12964-024-01946-7

Figure Lengend Snippet: Excessive accumulation of cytoplasmic mtDNA activates the cGAS-STING pathway to induce pyroptosis in cadmium-exposed AECs. MLE12 cells were treated with CdCl 2 for 24 h after VBIT4 intervention for 30 min. Cytosolic mtDNA ND1 and Dloop2 levels in MLE12 (A, n = 3). The expressions of IL-1β p17, GSDMD, cGAS, and STING protein and the phosphorylation level of TBK1 and NF-κB p65 in MLE12 cells were detected using western blot (B-C, n = 3). The fluorescence intensity of STING was detected by immunofluorescence (D, bar = 10 μm). Data are expressed as the mean ± SD. Differences among multiple groups were performed using ANOVA. Tukey’s test was used as a post hoc test for pairwise comparisons. * P < 0.05, ** P < 0.01, and *** P < 0.001

Article Snippet: Mouse alveolar epithelial cell lines MLE12 were purchased from ATCC (USA, CRL-2110).

Techniques: Phospho-proteomics, Western Blot, Fluorescence, Immunofluorescence

Journal: Cell Communication and Signaling : CCS

Article Title: Cadmium exposure triggers alveolar epithelial cell pyroptosis by inducing mitochondrial oxidative stress and activating the cGAS-STING pathway

doi: 10.1186/s12964-024-01946-7

Figure Lengend Snippet: Inhibition of the cGAS-STING pathway reduces lung injury and AEC pyroptosis induced by cadmium in mice. C57BL/6 J mice were given a daily intraperitoneal injection of CdCl 2 (3 mg/kg) and a daily intraperitoneal injection of RU.521 (5 mg/kg) (A). Weight change of mice (B, n = 7–10). The appearance of lung tissue (C). HE staining of lung tissue (D, bar = 100 μm). The inflammatory injury score was performed in a double-blind fashion (E, n = 6). The expressions of Nlrp3, pro-Il-1β, pro-Caspase1, Gsdmd, Sting, and Tbk1 mRNA in MLE12 cells were detected using Real-time PCR (F, n = 3). The expressions of NLRP3, Caspase-1 p20, IL-1β p17, GSDMD-N, cGAS, and STING protein and the phosphorylation level of TBK1 in the lung were detected using western blot (G-H, n = 3). The fluorescence intensity of SFTPC and GSDMD was detected by immunofluorescence (I, bar = 100 μm). Data are expressed as the mean ± SD. Differences among multiple groups were performed using ANOVA. Tukey’s test was used as a post hoc test for pairwise comparisons. * P < 0.05, ** P < 0.01, and *** P < 0.001

Article Snippet: Mouse alveolar epithelial cell lines MLE12 were purchased from ATCC (USA, CRL-2110).

Techniques: Inhibition, Injection, Staining, Real-time Polymerase Chain Reaction, Phospho-proteomics, Western Blot, Fluorescence, Immunofluorescence