Journal: Journal of Orthopaedic Translation

Article Title: Targeting the senescence-related genes MAPK12 and FOS to alleviate osteoarthritis

doi: 10.1016/j.jot.2024.06.008

Figure Lengend Snippet: Verification of mRNA and protein levels of candidate senescence-specific genes in osteoarthritis. (A) The mRNA levels of MAPK12, FOS, CYR61, TNFSF15 were investigated in human normal control and osteoarthritis (OA) cartilage tissue by qRT-PCR, n = 6. (B) HE, Safranine O staining and protein levels of MAPK12, FOS, CYR61, TNFSF15 were investigated in human normal control and osteoarthritis (OA) cartilage tissue by immunohistochemistry, n = 6. Scale bar = 100 μm. (C) The mRNA levels of MAPK12, FOS, CYR61, TNFSF15 were investigated in human C28/I2 chondrocytes treated with 10 ng/ml IL-1β for 48h by qRT-PCR, n = 3. (D) The protein levels of MAPK12, FOS, CYR61, TNFSF15 were investigated in human C28/I2 chondrocytes treated with 10 ng/ml IL-1β for 48h by western blotting, n = 3. Statistical analysis was conducted using unpaired Student's t test. P values were compared between OA group and normal control group, between IL-1β group and PBS control group. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, ns, no significance.

Article Snippet: The following primary antibodies were used: MMP13 (Proteintech, 18165-1-AP), Col2 (Proteintech, 28459-1-AP), p16 (Proteintech, 10883-1-AP), p21 (Proteintech, 10355-1-AP), ACAN (Affinity, DF7561), C-Myc-Tag (Affinity, T0052), GAPDH (Affinity, AF7021), PCNA (CST, 13110T), LMNB1 (Affinity, BF8009), ADAMTS5 (Affinity, DF13268), SOX9 (Santa Cruz, sc-166505), MAPK12 (Proteintech, 20184-1-AP), FOS (Proteintech, 66590-1-Ig), CYR61 (Proteintech, 67656-1-Ig) and TNFSF15 (Proteintech, 29899-1-AP).

Techniques: Control, Quantitative RT-PCR, Staining, Immunohistochemistry, Western Blot

Journal: Journal of Orthopaedic Translation

Article Title: Targeting the senescence-related genes MAPK12 and FOS to alleviate osteoarthritis

doi: 10.1016/j.jot.2024.06.008

Figure Lengend Snippet: MAPK12 over-expression inhibits senescence and DNA damage of C28/I2 cells induced by H 2 O 2 . (A) The protein levels of LMNB1, PCNA, p16, p21, MMP13 were investigated in C28/I2 cells by western blotting, n = 3. (B) EdU staining was adopted for cell proliferation capability of C28/I2 cells, n = 3. Scale bar = 100 μm. (C) The protein level of γ-H2AX was investigated in C28/I2 cells by immunofluorescence, n = 3. Scale bar = 100 μm. (D) SA-β-gal staining was adopted for senescence of C28/I2 cells, n = 3. Scale bar = 100 μm. Statistical analysis was conducted using one-way ANOVA. P values were compared between OE-MAPK12 group or control group and H 2 O 2 group. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, ns, no significance.

Article Snippet: The following primary antibodies were used: MMP13 (Proteintech, 18165-1-AP), Col2 (Proteintech, 28459-1-AP), p16 (Proteintech, 10883-1-AP), p21 (Proteintech, 10355-1-AP), ACAN (Affinity, DF7561), C-Myc-Tag (Affinity, T0052), GAPDH (Affinity, AF7021), PCNA (CST, 13110T), LMNB1 (Affinity, BF8009), ADAMTS5 (Affinity, DF13268), SOX9 (Santa Cruz, sc-166505), MAPK12 (Proteintech, 20184-1-AP), FOS (Proteintech, 66590-1-Ig), CYR61 (Proteintech, 67656-1-Ig) and TNFSF15 (Proteintech, 29899-1-AP).

Techniques: Over Expression, Western Blot, Staining, Immunofluorescence, Control

Journal: Journal of Orthopaedic Translation

Article Title: Targeting the senescence-related genes MAPK12 and FOS to alleviate osteoarthritis

doi: 10.1016/j.jot.2024.06.008

Figure Lengend Snippet: Over-expression of MAPK12 promotes cartilage anabolism and proliferation, inhibits cartilage catabolism and senescence of IL-1β stimulated C28/I2 cells. (A, B) The protein levels and quantification of LMNB1, PCNA, p16, p21, COL2, ACAN, SOX9, MMP13, ADAMTS5 were investigated in C28/I2 cells by western blotting, n = 3. (C, D) Representative image and quantification of EdU staining adopted for cell proliferation ability of C28/I2 cells, n = 3. Scale bar = 100 μm. Statistical analysis was conducted using one-way ANOVA. P values were compared between OE-MAPK12 group or control group and IL-1β group. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, ns, no significance.

Article Snippet: The following primary antibodies were used: MMP13 (Proteintech, 18165-1-AP), Col2 (Proteintech, 28459-1-AP), p16 (Proteintech, 10883-1-AP), p21 (Proteintech, 10355-1-AP), ACAN (Affinity, DF7561), C-Myc-Tag (Affinity, T0052), GAPDH (Affinity, AF7021), PCNA (CST, 13110T), LMNB1 (Affinity, BF8009), ADAMTS5 (Affinity, DF13268), SOX9 (Santa Cruz, sc-166505), MAPK12 (Proteintech, 20184-1-AP), FOS (Proteintech, 66590-1-Ig), CYR61 (Proteintech, 67656-1-Ig) and TNFSF15 (Proteintech, 29899-1-AP).

Techniques: Over Expression, Western Blot, Staining, Control

Journal: Journal of Orthopaedic Translation

Article Title: Targeting the senescence-related genes MAPK12 and FOS to alleviate osteoarthritis

doi: 10.1016/j.jot.2024.06.008

Figure Lengend Snippet: MAPK12 over-expression and FOS deficiency relieve osteoarthritis and joint pain in OA. (A) Representative images of HE, Safranine O staining and synovium in mice knee joint with or without MAPK12 over-expression or FOS knock-down adenovirus, n = 6. (B) OARSI score, meniscus total scores and synovitis score of mice knee joint with or without MAPK12 over-expression or FOS knock-down adenovirus, n = 6. (C, D) Representative images and quantification of Col2, ACAN, MMP13, Adamts5 protein levels in the knee cartilage of mice by immunohistochemistry, n = 6. Scale bar = 100 μm. (E) Paw withdrawl threshold of mice hind paw was measured using von frey filaments at 6 and 8 weeks post-DMM surgery, n = 6. (F) Response time of mice hind paw retraction or licking was recorded using hot plate at 6 and 8 weeks post-DMM surgery, n = 6. P values were compared between OE-MAPK12 and OE-control group, between siFOS group and si-control group, between OE-control or si-control group and sham group. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, ns, no significance.

Article Snippet: The following primary antibodies were used: MMP13 (Proteintech, 18165-1-AP), Col2 (Proteintech, 28459-1-AP), p16 (Proteintech, 10883-1-AP), p21 (Proteintech, 10355-1-AP), ACAN (Affinity, DF7561), C-Myc-Tag (Affinity, T0052), GAPDH (Affinity, AF7021), PCNA (CST, 13110T), LMNB1 (Affinity, BF8009), ADAMTS5 (Affinity, DF13268), SOX9 (Santa Cruz, sc-166505), MAPK12 (Proteintech, 20184-1-AP), FOS (Proteintech, 66590-1-Ig), CYR61 (Proteintech, 67656-1-Ig) and TNFSF15 (Proteintech, 29899-1-AP).

Techniques: Over Expression, Staining, Knockdown, Immunohistochemistry, Control

Journal: Journal of Orthopaedic Translation

Article Title: Targeting the senescence-related genes MAPK12 and FOS to alleviate osteoarthritis

doi: 10.1016/j.jot.2024.06.008

Figure Lengend Snippet: MAPK12 over-expression and FOS deficiency inhibit chondrocyte senescence in OA. (A) p16 protein levels were investigated in knee cartilage of mice by immunofluorescence. Scale bar = 50 μm. (B) p21 protein levels were investigated in knee cartilage of mice by immunofluorescence. Scale bar = 50 μm. (C) γ-H2AX protein levels were investigated in knee cartilage of mice by immunohistochemistry. Scale bar = 100 μm. (D) IL6 protein levels were investigated in knee cartilage of mice by immunohistochemistry. Scale bar = 100 μm. (E) Quantification of p16, p21, γ-H2AX and IL6 protein levels in knee cartilage of mice, n = 6. (F) Schematic illustration of the MAPK12 and FOS signaling pathways in OA by regulating cellular senescence. Statistical analysis was conducted using one-way ANOVA. P values were compared between OE-MAPK12 and OE-control group, between siFOS group and si-control group, between OE-control or si-control group and sham group. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, ns, no significance.

Article Snippet: The following primary antibodies were used: MMP13 (Proteintech, 18165-1-AP), Col2 (Proteintech, 28459-1-AP), p16 (Proteintech, 10883-1-AP), p21 (Proteintech, 10355-1-AP), ACAN (Affinity, DF7561), C-Myc-Tag (Affinity, T0052), GAPDH (Affinity, AF7021), PCNA (CST, 13110T), LMNB1 (Affinity, BF8009), ADAMTS5 (Affinity, DF13268), SOX9 (Santa Cruz, sc-166505), MAPK12 (Proteintech, 20184-1-AP), FOS (Proteintech, 66590-1-Ig), CYR61 (Proteintech, 67656-1-Ig) and TNFSF15 (Proteintech, 29899-1-AP).

Techniques: Over Expression, Immunofluorescence, Immunohistochemistry, Protein-Protein interactions, Control

Journal: AJNR. American journal of neuroradiology

Article Title: Macro- and Microstructural Changes in Patients with Spinocerebellar Ataxia Type 6: Assessment of Phylogenetic Subdivisions of the Cerebellum and the Brain Stem

doi: 10.3174/ajnr.A4085

Figure Lengend Snippet: Infratentorial parcellation map. The cerebellum is divided into 12 subdivisions, and the brain stem, into 6 subdivisions.

Article Snippet: Creation of the Infratentorial Parcellation Map We modified the Johns Hopkins University–Montreal Neurological Institute–based parcellation map in which the original left and right cerebellum were manually parcellated, and we divided the cerebellum into 12 subdivisions to create the infratentorial parcellation map as shown in .

Techniques:

Journal: AJNR. American journal of neuroradiology

Article Title: Macro- and Microstructural Changes in Patients with Spinocerebellar Ataxia Type 6: Assessment of Phylogenetic Subdivisions of the Cerebellum and the Brain Stem

doi: 10.3174/ajnr.A4085

Figure Lengend Snippet: Correlations between the data and ICARS scores in the infratentorial brain a

Article Snippet: Creation of the Infratentorial Parcellation Map We modified the Johns Hopkins University–Montreal Neurological Institute–based parcellation map in which the original left and right cerebellum were manually parcellated, and we divided the cerebellum into 12 subdivisions to create the infratentorial parcellation map as shown in .

Techniques: