Fig 2: (A) Representative images of HNSCC cell clusters stained for p16INK4a using immunofluorescence illustrate the HPV status of Cal-27 (HPV-negative) and UPCI-SCC 154 (HPV-positive) cells (scale bar, 40 µm). (B) Clonogenic survival assay of the HNSCC cell lines Cal-27 and UPCI-SCC 154 after 9 days. Data are presented as mean ± SD of 4 independent experiments. (C) Representative immunoblots demonstrate a rapid increase in cellular hnRNP K levels induced by IR (2 Gy), reaching maximum levels after 30-60 min before normalization of cellular hnRNP K levels within 24 h. Ratios represent hnRNP K/GAPDH referenced to non-irradiated control. (D) Dose-dependent accumulation of cellular hnRNP K 1 h after IR. (E) Immunofluorescence microscopy indicated cytoplasmic hnRNP K accumulation 1 h after irradiating cells with 2 Gy (scale bar, 20 µm). HNSCC, head and neck squamous cell carcinoma; hnRNP K, heterogenous nuclear ribonucleoprotein K; HPV, human papillomavirus; IR, ionizing radiation.

Fig 3: Hypoxia-induced cleavage and nuclear localization of DLK1 in GBM cells.a Representative images and densitometric analysis of western blots showing DLK1 expression and cleavage in different mouse (PIGPC) and human (U3046MG, U3035MG, U3082MG, U3084MG, U3065MG, T98G, and U251MG) GBM cell lines. Cells were grown at 21% or 1% O2, as indicated, for 48 h. SDHA was used as loading control. b Representative images and densitometric analysis of western blots showing nuclear localization of a DLK1 intracellular fragment. Cellular fractionation experiments were performed in U3082MG grown at 21% or 1% O2 for 48 h. GAPDH and TBP were used as cytoplasmic and nuclear fractions controls, respectively. c Representative images and relative quantifications of immunofluorescent stainings showing DLK1 localization in U3082MG cells grown at 21% or 1% O2 for 48 h. Statistical analysis: independent experimental replicates are as follow, a for PIGPC, U3046MG, U3082MG, U3084MG, and U3065MG n = 4, for U035MG, U251MG, and T98G n = 3; in b n = 3 and in c n = 8. All data are expressed as mean ± SEM, statistical significance was determined by Mann–Whitney test applied to a and t-test with Welch’s correction applied to b and c. In the whole figure significance is represented as *p < 0.05, **p < 0.01, and ***p < 0.001 vs. respective 21% O2 controls.

Fig 4: Direct regulation of SCIN by miR-301a-5p in gastric cancer. (a) SCIN was predicted as target as miR-301a-5p via bioinformatic analyses. (b) Expression level of SCIN in TCGA database. (c-d) mRNA and protein expression level of SCIN in GC cell lines by qPCR and western blot. GAPDH was used as internal control. (e-f) SCIN mRNA and protein level of GC patient sample was determined by qRCR and western blot. (g) After transfecting mimics of miR-301a-5p in cell lines, the alteration of SCIN mRNA and protein were measured by qPCR and western blot. GAPDH was used as internal control. (h) After transfecting inhibitor of miR-301a-5p in cell lines, the alteration of SCIN mRNA and protein were measured by qPCR and western blot. GAPDH was used as internal control. (i) miR-301a-5p binding sites in the 3'-UTR of SCIN mRNA. Dual luciferase reports assays exhibited that miR-301a-5p markedly suppressed luciferase activity of HEK-293T carrying wild-type reporter constructs. * p<0.05, ** p<0.01, *** p<0.001, **** p<0.0001.

Fig 5: Dynamic expression of Kindlin-1 in skin wound healing in mice. Assessment of Kindlin-1 in skin wound healing in mice with or without EGF treatment. (A1)Construction of wound healing model in mice. (A2) Healing rate of EGF(-) and EGF(+) group at the 14th day. *P<0.05. (B1) Kindlin-1 expression was detected by real-time PCR. (C1) Kindlin-1 expression was detected by western blot analysis and with GAPDH as an internal control. (C2) Kindlin-1 expression was quantified and presented by the fold changes compared with the relative control [value of EGF(-) 0 days or EGF(+) 0 days]. *P<0.05, significant difference compared to the normal group [EGF(-) 0 days, EGF(+) 0 days]. #P<0.05.