Fig 1: MRPS16 knockdown inhibits tumor growth. A. Representative images of the brain orthotopic tumor by U-138MG/Luc and U-87MG/Luc cells injected into the brain parenchyma of nude mice (left panels) and histogram analysis of luminescence representing the size of the tumors measured after four weeks (right panels). n = 5. B. Representative images (left panels) of IF staining with Ki-67 and histogram analysis of the percentage of Ki-67 (+) cells (right panels). C. Representative images of IHC staining with MRPS16 and Snail. Statistical significance was assessed using one-way ANOVA followed by Dunnett's tests for multiple comparisons. Scale bars: 50μm. *P < 0.05, **P < 0.01 and ***P < 0.001. None: non infected cells.
Fig 2: MRPS16 over-expression increases Snail via PI3K/AKT pathway and binds with each other. A. U-138MG and U-87MG cells transfected with MRPS16 plasmid and co-cultured with PI3K/AKT signaling inhibitors were reaped, and the lysates were immune-blotted for P-p85, Tot.p85 and β-actin. B. U-138MG and U-87MG cells transfected with MRPS16 plasmid and co-cultured with PI3K/AKT signaling inhibitors were reaped, and the lysates were immune-blotted for p-AKTSer473, AKT and β-actin. C. U-138MG and U-87MG cells transfected with MRPS16 plasmid and co-cultured with PI3K/AKT signaling inhibitors were reaped, and the lysates were immune-blotted for Snail, Slug and GAPDH. D. Co-IP experiment indicated that MRPS16 could bind with Snail. Statistical significance was tested using one-way ANOVA (Dunnett's tests) for multiple comparison and two-tailed t-tests. * P < 0.05 and **P < 0.01.
Fig 3: MRPS16 knockdown suppresses U-87MG cells progression. A-B. Validation of knocking down MRPS16 in U-87MG cells by Western blot and qRT-PCR. C. Growth curves between sh-NC, sh-MRPS16#1 and sh-MRPS16#2 groups by CCK-8 assay. D. Knockdown of MRPS16 suppressed U-87MG cell proliferation. Percentage of EdU (+) is expressed in the panel. E. Knockdown of MRPS16 suppressed colony formation and histogram quantification (panels). F - G. Transwell migration and invasion assay showing that knockdown of MRPS16 suppressed cell migration and invasion. The numbers of migrating and invading cells are measured in the panel. Bars: 50μm. Statistical significance was tested using one-way ANOVA (Dunnett's tests) for multiple comparison and two-tailed t-tests. All the results are shown as the Mean ± Standard Deviation (SD) of three independent repeated experiments at least. *P < 0.05, **P < 0.01 and ***P < 0.001. None: non infected cells.
Fig 4: MRPS16 over-expression facilitates U-138MG cells progression. A-B. MRPS16 protein and mRNA expression levels by Western blot and qRT-PCR in NHA, HA, U-138MG, U-251MG and U-87MG. C - D. The over-expression efficiency for MRPS16 was verified in U-138MG by Western blot and qRT-PCR. E. Growth curves between None, Vector and MRPS16 (OE) by CCK-8 assay. F. MRPS16 over-expression promoted U-138MG cell proliferation. Percentage of EdU (+) is expressed in the panel. G. Over-expression of MRPS16 obviously facilitated colony formation and histogram quantification (panels). H-I. Transwell migration and invasion assay showing that over-expression of MRPS16 facilitated cell migration and invasion. The numbers of migrating and invading cells are measured in the panel. Bars: 50μm. Statistical significance was tested using one-way ANOVA (Dunnett's tests) for multiple comparison and two-tailed t-tests. All the results are shown as the Mean ± Standard Deviation (SD) of three independent repeated experiments at least. *P < 0.05, **P < 0.01 and ***P < 0.001. None: non infected cells.
Fig 5: MRPS16 promotes tumor progression by the PI3K/AKT/Snail axis. A. Knockdown of MRPS16 decreased the protein levels of p-PI3K, p-AKT and Snail. B-C. Validation of over-expressing Snail in U-138MG and U-87MG cells by Western blot and qRT-PCR. D. Growth curves were tested by CCK-8 assay. E. Snail over-expression rescued the effects of MRPS16 knockdown in suppressing tumor cells growth. F. Over-expression of Snail rescued the effects of knockdown of MRPS16 in suppressing tumor cells colony formation. G - H. Transwell migration and invasion experiment was adopted as mentioned above. The numbers of migrating and invading cells are measured in the panel. Bars: 50μm. Statistical significance was tested using one-way ANOVA (Dunnett's tests) for multiple comparison and two-tailed t-tests. All the results are shown as the Mean ± Standard Deviation (SD) of three independent repeated experiments at least. **P < 0.01 and ***P < 0.001. None: non infected cells.
Supplier Page from Abcam for Anti-MRPS16 antibody [EPR8835]