Journal: Cell death & disease

Article Title: Deletion of Mettl3 in mesenchymal stem cells promotes acute myeloid leukemia resistance to chemotherapy.

doi: 10.1038/s41419-023-06325-7

Figure Lengend Snippet: Fig. 1 Deletion of Mettl3 in MSCs induces high marrow adiposity in mice. A Western blot analysis of METTL3 in BMMSCs isolated from mice. B Detections of m6A levels in total RNA in BMMSCs from mice using EpiQuik m6A RNA Methylation Quantification Kits. C Representative images of H&E staining and FABP4 immunohistochemical staining of the distal femora from mice. Scale Bars: 100 μm. D Quantification of adipocyte number and area per tissue area in the distal marrow (n = 8). E ELISA examined the contents of supernatant adipokines in bone marrow rinses of mice (n = 6). F Representative images and quantitative analyses of Alizarin Red Solution (ARS) staining of BMMSCs isolated from mice. Scale Bars: 200 μm. G Representative images and quantitative analyses of ORO staining of BMMSCs isolated from mice. Scale Bars: 200 μm. The data are presented as mean ± SD. The two-tailed unpaired Student’s t test was used to compare two groups and one-way ANOVA with Dunnett’s multiple comparisons test was used for data with more than two groups. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, ns not significant.

Article Snippet: In the supernatant, based on the Cell Death and Disease (2023) 14:796 manufacturer’s directions, the levels of adipokines, like resistin, leptin, and growth hormone, were quantified employing the kits of Mouse Resistin ELISA (Cat# CSB-E06886m, CUSABIO, Wuhan, Hubei, China), Mouse Leptin ELISA (Cat# CSB-E04650m, CUSABIO), and Mouse Growth Hormone ELISA (Cat# CSB-E07343m, CUSABIO), respectively.

Techniques: Western Blot, Isolation, Methylation, Staining, Immunohistochemical staining, Enzyme-linked Immunosorbent Assay, Two Tailed Test

Journal: Cell death & disease

Article Title: Deletion of Mettl3 in mesenchymal stem cells promotes acute myeloid leukemia resistance to chemotherapy.

doi: 10.1038/s41419-023-06325-7

Figure Lengend Snippet: Fig. 4 METTL3 deletion reduces AML chemosensitivity by inducing adipogenic differentiation of OP9 cells. A Representative images and quantification of ORO staining in Mettl3 overexpression cells after 14 days of adipogenic induction. Scale bar, 100 µm. B qPCR analysis of the expression of adipogenic marker genes, Adipoq, Cebpa, Lpl, Plin1, CD36, and Pparγ in Mettl3 overexpression cells under adipogenic conditions. C ORO staining of Mettl3 knockdown cells following adipogenic induction. Scale bar, 100 µm. Absorbance at OD500 was determined for ORO staining in isopropanol at room temperature. D qPCR analysis of adipogenic lineage–associated gene expression after induction of differentiation in Mettl3 knockdown cells. E ELISA detection of adipokine concentration in the culture medium before induction of differentiation and on day 14 of Mettl3 overexpression cells. F ELISA detection of adipokine concentration in the culture medium before induction of differentiation and on day 14 of Mettl3 knockdown cells. G After adipogenic differentiation of Mettl3 overexpression cells, AML cells were co-cultured with them for 24 h to test their chemoresistance to Ara-C and DNR. H After adipogenic differentiation of Mettl3 knockdown cells, AML cells were co-cultured with them for 24 h to test their chemoresistance to Ara-C and DNR. Blank, blank control; OE-NC, negative control of overexpression; Mettl3-OE, overexpression of mouse Mettl3; sh-NC, negative control of knockdown; shMettl3-1 and shMettl3-2, independent shRNAs targeting mouse Mettl3. The data are presented as mean ± SD. The two-tailed unpaired Student’s t test was used to compare two groups and one-way ANOVA with Dunnett’s multiple comparisons test was performed for multiple groups. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, ns not significant.

Article Snippet: In the supernatant, based on the Cell Death and Disease (2023) 14:796 manufacturer’s directions, the levels of adipokines, like resistin, leptin, and growth hormone, were quantified employing the kits of Mouse Resistin ELISA (Cat# CSB-E06886m, CUSABIO, Wuhan, Hubei, China), Mouse Leptin ELISA (Cat# CSB-E04650m, CUSABIO), and Mouse Growth Hormone ELISA (Cat# CSB-E07343m, CUSABIO), respectively.

Techniques: Staining, Over Expression, Expressing, Marker, Knockdown, Gene Expression, Enzyme-linked Immunosorbent Assay, Concentration Assay, Cell Culture, Control, Negative Control, Two Tailed Test

Journal: Cell death & disease

Article Title: Deletion of Mettl3 in mesenchymal stem cells promotes acute myeloid leukemia resistance to chemotherapy.

doi: 10.1038/s41419-023-06325-7

Figure Lengend Snippet: Fig. 6 AKT inhibitor MK2206 mediates the adipogenesis level of OP9 cells. A Western blot analysis of AKT and p-AKT1 (Ser473) in OP9 cells treated with MK2206, an AKT inhibitor. B ORO staining of DMSO and MK2206 treated cells after adipo-induced for 14 days. Scale bar, 100 μm. C ELISA detected the concentration of adipokines in the culture medium before induction of differentiation and on day 14 under treatment with MK2206. D After being treated with MK2206 for 18 h and adipo-induced for 14 days, co-culturing with AML cells for 24 h was performed to test the chemoresistance of AML cells to Ara-C and DNR. The data are presented as mean ± SD. The two-tailed unpaired Student’s t test was used to compare two groups and one-way ANOVA with Dunnett’s multiple comparisons test was performed for multiple groups. *P < 0.05, **P < 0.01, ***P < 0.001, ns not significant.

Article Snippet: In the supernatant, based on the Cell Death and Disease (2023) 14:796 manufacturer’s directions, the levels of adipokines, like resistin, leptin, and growth hormone, were quantified employing the kits of Mouse Resistin ELISA (Cat# CSB-E06886m, CUSABIO, Wuhan, Hubei, China), Mouse Leptin ELISA (Cat# CSB-E04650m, CUSABIO), and Mouse Growth Hormone ELISA (Cat# CSB-E07343m, CUSABIO), respectively.

Techniques: Western Blot, Staining, Enzyme-linked Immunosorbent Assay, Concentration Assay, Two Tailed Test