The authors of the original article “miR- 210 transferred by lung cancer cell-derived exosomes may act as proangiogenic factor in cancer-associated fibroblasts by modulating JAK2/STAT3 pathway” (Clinical Science (2020) 134(7), DOI: 10.1042/CS20200039) have acknowledged an error regarding image duplication in Figures 3, 5 and 7 of their published paper. The authors would like to apologise for this error. The corrected Figures 3, 5 and 7 are provided below and the author has additionally provided the raw data to support this as a Supplementary File. This correction does not modify data interpretation of the original article.
Regulatory effects of the LC cell-derived exosomes on the proangiogenic switch of CAFs through modulating the JAK2/STAT3 signaling pathway
(A) Increased phosphorylation of JAK2 and STAT3 induced by the administration of LC cell-derived exosomes in NIH/3T3 cells after stimulation of LC cell-derived exosome. Western blot analysis revealing the expression of nuclear P-STAT3 (nuclear protein marker: Histone H3). (B) Increased accumulation of P-STAT3 in the nucleus of the NIH/3T3 cells induced by LC exosomes according to confocal microscope images. (C) Decreased expression of the mRNA of the FGF2, MMP9 and VEGFa in CAFs after the administration of Stattic; *P<0.05, **P<0.01. (D) Decreased expressions of VEGFa, MMP9, FGF2 and phosphorylation of STAT3 and JAK2 induced by Stattic which is measured by Western blot assay in CAF. (E,F) Promoted MS-1 cell proliferation induced by the CM of NIH/3T3-cell that treated with LC cell-derived exosomes could be blocked by Stattic. *P<0.05, **P<0.01. (G,H) Transwell migration assay and tube formation indicated that elevated MS-1 cell migration and tube formation induced by the CM-NIH/3T3-cell that treated with LC cell-derived exosomes were weakened by Stattic. *P<0.05, **P<0.01.
(A) Increased phosphorylation of JAK2 and STAT3 induced by the administration of LC cell-derived exosomes in NIH/3T3 cells after stimulation of LC cell-derived exosome. Western blot analysis revealing the expression of nuclear P-STAT3 (nuclear protein marker: Histone H3). (B) Increased accumulation of P-STAT3 in the nucleus of the NIH/3T3 cells induced by LC exosomes according to confocal microscope images. (C) Decreased expression of the mRNA of the FGF2, MMP9 and VEGFa in CAFs after the administration of Stattic; *P<0.05, **P<0.01. (D) Decreased expressions of VEGFa, MMP9, FGF2 and phosphorylation of STAT3 and JAK2 induced by Stattic which is measured by Western blot assay in CAF. (E,F) Promoted MS-1 cell proliferation induced by the CM of NIH/3T3-cell that treated with LC cell-derived exosomes could be blocked by Stattic. *P<0.05, **P<0.01. (G,H) Transwell migration assay and tube formation indicated that elevated MS-1 cell migration and tube formation induced by the CM-NIH/3T3-cell that treated with LC cell-derived exosomes were weakened by Stattic. *P<0.05, **P<0.01.
Regulatory effects of the miR-210 regulates on the proangiogenic switch of CAFs
(A) Increased expression levels of proangiogenic factors and phosphorylation of STAT3 and JAK2 induced by the exosomes from miR-210-mimic-transfected LC cells, and inhibitory effects induced by the exosomes from anti-miR-210-transfected cells in NIH/3T3 cells according to the Western blot analysis. (B-E) The promotion effect of CM from NIH/3T3 cell that stimulated by the exosomes from miR- 210-mimic-transfected A549 cells and the inhibitory effect of CM from NIH/3T3 cell that stimulated by the exosomes from anti-miR-210-transfected H460 cells on MS-1 cell proliferation according to the CCK-8 and colony formation assay; *P<0.05, **P<0.01 vs. the CM from untreated NIH/3T3 cells. (F-I) Increased MS-1 cell migration (transwell migration assay) and tube formation induced by the CM from NIH/3T3 cells that treated with miR-210-mimic-transfected A549-secreted exosomes on MS-1 cells and the reduction effect of CM from NIH/3T3 cells that treated with anti-miR-210-transfected H460-secreted exosomes on MS-1 cells; *P<0.05, **P<0.01.
(A) Increased expression levels of proangiogenic factors and phosphorylation of STAT3 and JAK2 induced by the exosomes from miR-210-mimic-transfected LC cells, and inhibitory effects induced by the exosomes from anti-miR-210-transfected cells in NIH/3T3 cells according to the Western blot analysis. (B-E) The promotion effect of CM from NIH/3T3 cell that stimulated by the exosomes from miR- 210-mimic-transfected A549 cells and the inhibitory effect of CM from NIH/3T3 cell that stimulated by the exosomes from anti-miR-210-transfected H460 cells on MS-1 cell proliferation according to the CCK-8 and colony formation assay; *P<0.05, **P<0.01 vs. the CM from untreated NIH/3T3 cells. (F-I) Increased MS-1 cell migration (transwell migration assay) and tube formation induced by the CM from NIH/3T3 cells that treated with miR-210-mimic-transfected A549-secreted exosomes on MS-1 cells and the reduction effect of CM from NIH/3T3 cells that treated with anti-miR-210-transfected H460-secreted exosomes on MS-1 cells; *P<0.05, **P<0.01.
TET2 was identified as the target of exosomal miR-210 in CAFs
(A) RNA pull-down was performed to identify potential target of miR-210, which is plotted as a flow diagram. The biotin-coupled RNA complex was pulled down and then subjected to RNA-Sequencing. Abundance of Target gene in bound fractions was analyzed by RT-qPCR. (B) The compatible result of RNA-Sequencing in two exosomes was shown in the heatmap. (C) A schematic diagram showed miR-210 binding sites in the 3′-UTR of TET2. (D) RT-PCR assay showed expression of TET2 in Biotin-coupled RNA complex. (E) The wild-type and mutant of miR-210 binding sites in the 3′-UTR of TET2 were shown in a schematic diagram. The Dual-Luciferase reporter assay was used in NIH/3T3 cells. Luciferase activity was determined by Glomax 96 luminometer at 48 h. (F) Increased expression levels of proangiogenic factors and phosphorylation of STAT3 and JAK2 and reduced expression levels of TET2 induced by the exosomes from miR-210-mimic-transfected LC cells could be neglected by transfection of Lv-TET2 in NIH/3T3 cells according to the Western blot analysis and vice versa. (G-J) The promoting effect of CM from NIH/3T3 cells that stimulated by the exosomes from miR-210-mimic-transfected A549 cells on MS-1 cell proliferation could be neutralized by simultaneous transfection of Lv-TET2 in NIH/3T3 cells according to the CCK-8 and colony formation assay and vice versa. *P<0.05, **P<0.01.
(A) RNA pull-down was performed to identify potential target of miR-210, which is plotted as a flow diagram. The biotin-coupled RNA complex was pulled down and then subjected to RNA-Sequencing. Abundance of Target gene in bound fractions was analyzed by RT-qPCR. (B) The compatible result of RNA-Sequencing in two exosomes was shown in the heatmap. (C) A schematic diagram showed miR-210 binding sites in the 3′-UTR of TET2. (D) RT-PCR assay showed expression of TET2 in Biotin-coupled RNA complex. (E) The wild-type and mutant of miR-210 binding sites in the 3′-UTR of TET2 were shown in a schematic diagram. The Dual-Luciferase reporter assay was used in NIH/3T3 cells. Luciferase activity was determined by Glomax 96 luminometer at 48 h. (F) Increased expression levels of proangiogenic factors and phosphorylation of STAT3 and JAK2 and reduced expression levels of TET2 induced by the exosomes from miR-210-mimic-transfected LC cells could be neglected by transfection of Lv-TET2 in NIH/3T3 cells according to the Western blot analysis and vice versa. (G-J) The promoting effect of CM from NIH/3T3 cells that stimulated by the exosomes from miR-210-mimic-transfected A549 cells on MS-1 cell proliferation could be neutralized by simultaneous transfection of Lv-TET2 in NIH/3T3 cells according to the CCK-8 and colony formation assay and vice versa. *P<0.05, **P<0.01.
