Fig 2: PAR2 antagonist AZ3451 suppresses IL-1β-induced inflammation response, cartilage degradation and premature senescence in chondrocytes. (A) Chemical structure of AZ3451. (B, C) Representative western blots and quantification data of PAR2 in chondrocytes of each group. (D, E) Representative western blots and quantification data of iNOS, COX2, MMP1, MMP13 and ADAMTS5 in chondrocytes of each group. (F, G) Representative western blots and quantification data of PAR2, Aggrecan, Collagen II and Sox9 in chondrocytes of each group. (H, I) Representative western blots and quantification data of p16INK4a in chondrocytes of each group. (J, K) SA-β gal staining assay was performed in chondrocytes as treated above. Data are shown as the mean ± SD. Significant differences between groups are indicated as ***P <0.001, **P < 0.01 and *P < 0.05.

Fig 3: 5-hydroxytryptamine receptor 7 (HTR7) knockdown inhibits laryngeal cancer proliferation. (A) Western blotting analysis of cyclin D1, cyclin E, p16, and p21 levels when HTR7 was knocked down in TU212 cells. GAPDH was used as the loading control. (B) 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay of the effect of HTR7 knockdown on laryngeal cancer proliferation. (C) Colony formation assay of the effect of HTR7 knockdown on laryngeal cancer proliferation using crystal violet staining. Microscope magnification 200×. (D) BrdU incorporation assay of the effect of HTR7 knockdown on laryngeal cancer proliferation. Error bars represent the mean ± STDEV. *P<0.05.

Fig 4: HuR regulates autophagy activation in high glucose‐treated NP cells. The cells were transfected with LV‐NC or LV‐shHuR or LV‐HuR before high‐glucose treatment (100 mM). A, TEM images of autophagic vesicles in rat NP cells (bar: 0.5 μm). B, The LC3‐II was detected by immunofluorescence combined with DAPI staining for nuclei (scale bar: 20 μm). C‐E, The protein expression of p62 and the ratio of LC3‐II/ LC3‐I in NP cells treated with or without high‐glucose treatment (100 mM) for 6 hours and transfected with lentivirus, detected by Western blot and quantified by Image J software. F, EdU staining for NP cells (scale bar: 50 μm). G, SA‐β‐gal staining for NP cells (scale bar: 50 μm). H, I, The protein expression of p21 and p16INK4a in NP cells with or without rapamycin (the classical autophagy activator, 10 μM, 24 hours), detected by Western blot. All data were shown as mean ± SD. *P < .05, **P < .01

Fig 5: Overexpression of Atg7, instead of HuR, ameliorates the progress of DB‐IVDD in vivo. A, T2 weighted MRI of a rat‐tail disc at 3 weeks after disc puncture surgery, with or without lentivirus transfection (white arrows). B, The respective Pfirrmann grade scores of a rat‐tail disc. C, X‐ray of a rat‐tail disc at 4 weeks after disc puncture surgery in normal and diabetic rats, with or without lentivirus transfection (white arrows). D, The disc height index (DHI) of a rat‐tail disc. E, Representative H‐E, S‐O and Alcian Blue staining of NP tissues (scale bar: 800 μm). F, The histological grades evaluated in three groups. G‐I, The immunohistochemical staining of p16 and LC3‐II in intervertebral disc sections; the levels were determined using Image‐Pro Plus software. All data were shown as mean ± SD. *P < .05, **P < .01