Fig 2: Overexpression of miR-873 significantly inhibits BMSC proliferation. (A) The transfection efficiencies of miR-873 inhibitors and mimics were confirmed by RT-qPCR. (B) The proliferation of BMSCs was detected by Cell Counting Kit-8 assay. (C) RT-qPCR and western blotting were used to measure the relative expression levels of PCNA mRNA and protein, respectively. (D) Flow cytometry was used to evaluate the proportion of BMSCs at different phases. (E and F) The DNA contents in miR-NC and miR-873 mimics transfected cells were evaluated. (G and I) The differentiation of cells was inhibited by the transfection with miR-873 mimics (magnification, ×200). (H and J) Western blots revealed the expression of differentiation-related markers were reduced in miR-873 mimic transfected cells. The data are presented as the mean ± SD; *P<0.05 vs. miR-NC. Bglap2, bone γ-carboxyglutamate protein 2; BMSCs, bone marrow mesenchymal stem cells; C/EBPα, CCATT/enhancer binding protein α; miR, microRNA; NC, negative control; OD, optical density; PCNA, proliferating cell nuclear antigen; PPARγ, peroxisome proliferator-activated receptor γ; RT-qPCR, reverse transcription-quantitative PCR; RunX, runt-related transcription factor; SREBP1, sterol regulatory element binding protein 1.

Fig 3: TPX2 knockdown suppresses the proliferation of breast cancer cells by downregulating NCOA5. (A) MCF7 cells were transfected with Ov-NCOA5 or Ov-NC. The transfection efficiency of si-NCOA5 in MCF7 cells was validated via RT-qPCR. ***P<0.001. (B) MCF7 cells were transfected with si-TPX2 or co-transfected with si-TPX2 and Ov-NCOA5. Cell Counting Kit-8 assay for determination of the viability of MCF7 cells. ***P<0.001 vs. si-TPX2. ##P<0.01, ###P<0.001 vs. si-TPX2 + Ov-NC. (C) MCF7 cells were transfected with si-TPX2 or co-transfected with si-TPX2 and Ov-NCOA5. Western blot analysis for determination of Ki67 and PCNA protein expression in MCF7 cells. **P<0.01, ***P<0.001. NC, negative control; NCOA5, nuclear receptor coactivator 5; OD, optical density; Ov, overexpression plasmid; PCNA, proliferating cell nuclear antigen; RT-qPCR, reverse transcription-quantitative PCR; si, small interfering RNA; TPX2, targeting protein for xenopus kinesin-like protein 2.

Fig 4: Representative images of immunohistochemical staining.A few PCNA-positive cells are shown in adenoma of the lung (A) Many PCNA-positive cells were found in lung adenocarcinoma (B), thymic malignant lymphoma (E), and thymic malignant lymphoma (TML) metastasis in the lungs (F). A few SPC-positive cells indicate an adenoma of the lungs (C), but many SPC-positive cells indicate adenocarcinoma of the lung (D). No CC10-positive cells were observed in the adenoma of the lungs (G), but CC10-positive cells were present in the bronchial epithelium at the center of the TML lung metastasis. Scale bar represents 100 μm.

Fig 5: Deficiency of Parp16 suppresses PDGF-BB-induced rVSMC proliferation and migration through ER stress. Following transfection with control siRNA (siCTL) or Parp16 siRNA (siPARP16) for 72 h, rVSMCs were treated with 20 ng/mL PDGF for 24 h, cell extracts were collected for determining the protein levels of PARP16, PCNA, MMP9, cyclin D1, and cyclin E by Western blot (A); cell lysates were immunoblotted with p-PERK, p-eIF2α, BIP, p-IRE1α, spliced XBP-1 and calnexin antibodies (B). rVSMCs were transfected with Parp16 siRNA (siPARP16) or control siRNA (siCTL) for 72 h, and then stimulated with 20 ng/mL PDGF-BB for 24 or 48 h, the EdU positive cells were showed green color and cell nuclei stained with DAPI showed blue color, scale bars: 200 μm (C); the migratory ability of rVSMCs was determined by Transwell assay, scale bars: 200 μm (D) and wound healing assay, scale bars: 400 μm (E). All data are represented as means ± SEM; ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001 vs. control; &&P < 0.01, &&&P < 0.001 vs. PDGF + siCTL, each acquired from three individual experiments.