Fig 2: Contribution of LUC7L3 to spindle assembly through promoting spindle-associated protein translationA. Observation of cell multinucleation in LUC7L3-knockdown and GFP-RNase H1-rescued A549 and HeLa cells. Statistical analysis of the number of multinucleated cells was shown. n＞100.B. Immunofluorescence analysis of mitotic spindles in LUC7L3-depleted HeLa cells. Red fluorescence indicates alpha-tubulin, green fluorescence indicates pericentrin, and blue indicates the nucleus stained with DAPI.C. Immunofluorescence analysis of mitotic spindles in LUC7L3-depleted and SFB-LUC7L3-rescued HeLa cells. Statistical analysis of the proportion of cells with multipolar spindles was shown. n＞100.D. Western blot analysis of protein levels of LUC7L3, KIF2A, NDC80, CEP70, and CEP170 in LUC7L3-depleted A549 cells. The protein level was analyzed in asynchronized cells and nocodazole-synchronized mitotic cells. Statistical quantification of each protein was shown. Asyn: Asynchronous.E. Immunofluorescence analysis of intracellular CEP170 fluorescence in LUC7L3-depleted HeLa cells. Statistical analysis of CEP170 fluorescence signals were shown. n＞40. a. u. means arbitrary units.F. Ribosomal fraction isolation assay in A549 cells overexpressing SFB-LUC7L3 or with LUC7L3 depletion. The ratio of indicated mRNA in polysome fractions to sub-polysome fractions was explored using qPCR. Statistical analysis of the ratio for each gene was shown.G. Western blot analysis of protein levels of KIF2A, NDC80, CEP70, and CEP170 in A549 ​cells overexpressing SFB-SRSF1. Statistical quantification of each protein was shown. ∗p ​< ​0.05, ∗∗p ​< ​0.01, ∗∗∗p ​< ​0.001, ∗∗∗∗p ​< ​0.0001, indicating statistical significance and ns indicates non-significance. Scale bar: 10 ​μm. Dash lines outlined cell shape.

Fig 3: Mechanistic graph summarizing a possible regulatory role of circ_000829 in RCC. Circ_000829 inhibits the alternative splicing of SLC39A14 mRNA by binding to SRSF1, thereby inhibiting the proliferation of RCC cells in vitro and tumorigenesis in vivo.

Fig 4: The gene CCL21 induces the expression of MALAT1, and MALAT1 further increases the SRSF1 expression to activate the mTOR pathway, thereby promoting the migration as well as EMT of GC cells and ultimately contributing to the development of GC

Fig 5: Ectopic MYC activation altered pre‐mRNA splicing without transcriptional regulation of CLK AMYC induction increased expression of SRSF1 and PRMT5, but not CLK family kinases. The same samples of Fig 5A were analyzed with RT–PCR.BChanges in CPM values of CLK families, PRMT5, and other splicing‐related genes calculated from RNA‐Seqs data of MYC‐inducible SK‐MEl‐28 cells treated with Dox for 54 h.CThe numbers of AS events modulated by Dox with a BF > 20 and |ΔPSI| > 0.1 were counted and categorized according to the AS type.DComparison between MYC induction‐dependent and CLK inhibitor‐dependent alternative splicing events.EEffects of T‐025 on MYC‐inducible SK‐MEL‐28 cells were analyzed by immunoblotting. SK‐MEl‐28 cells pretreated with Dox for 48 h were additionally treated with T‐025 for 24 h.F, GExon 3 skipping of NOP16 was induced by T‐25. MYC‐inducible SK‐MEL‐28 cells pretreated with Dox were treated with 100 nmol/l T‐025 for 6 h. NOP16 exon 2–3 and 2–4 transcripts were measured by quantitative RT–PCR, and the PSI value of each sample was calculated (F). The copies of exon 2–3 transcripts are shown (G).Data information: In (A), an unpaired Student's t‐test was performed. In (F and G), Tukey's test was performed. In (A, F, and G), data are shown as the means ± s.d. of three independent experiments (n = 3).Source data are available online for this figure.