Additional file 1 of LncRNA AC006064.4–201 serves as a novel molecular marker in alleviating cartilage senescence and protecting against osteoarthritis by destabilizing CDKN1B mRNA via interacting with PTBP1
Creators: name: Shen, Panyang
name: Gao, Jun
name: Huang, Shaohan
name: You, Chenan
name: Wang, Haitao
name: Chen, Pengyu
name: Yao, Teng
name: Gao, Tianyou
name: Zhou, Bohao
name: Shen, Shuying
name: Zhao, Xing
name: Ma, Jianjun
Publisher: name: figshare
Year: 2023
Subjects: subject: Biophysics
subject: Biochemistry
subject: Medicine
subject: Cell Biology
subject: Molecular Biology
subject: Physiology
subject: FOS: Biological sciences
subject: Biological Sciences not elsewhere classified
subject: Developmental Biology
subject: Cancer

description: Additional file 1: Supplementary Figure 1. (A) Expressionof ten lncRNAs in HCs assessed by qRT-PCR after treating with 10ng/ml IL-1β. (n=9, 3 donors for three replicates) * p&lt;0.05, ** p&lt;0.01. (B) Fluorescence intensity of FISH staining forAC006064.4-201 and IF staining of p16INK4a and Mmp3. * p&lt;0.05, ** p&lt;0.01,*** p&lt;0.001. (C) Expressionof AC006064.4-201 in HCs after cheating with different concentrations ofDoxorubicin (0 nm/ml, 100nm/ml and 200nm/ml). (n=9, 3 donors for three replicates) * p&lt;0.05. Supplementary Figure 2. (A) QRT-PCR ofAC006064.4-201 in HCs when AC006064.4-201 was knocked down. (n=9, 3 donors for three replicates) *** p&lt;0.001. (B) QRT-PCR ofAC006064.4-201 in HCs when AC006064.4-201 was overexpressed. (n=9, 3 donors for three replicates) ** p&lt;0.01. (C) Western blot ofYBX3, APOBEC3C, PTBP1, LIN28A and IMPDH1 when they were separately knockeddown. (D) Knock down efficiency of CDKN1B shRNAs assessed by qRT-PCR. (n=9, 3 donors for three replicates) *** p&lt;0.001. (E) MRNA levels ofMmp3, Mmp13, Sox9, Aggrecan, p16INK4a, p21 and p53 assessed by qRT-PCR whenCDKN1B was knocked down in HCs. (n=9,3 donors for three replicates) * p&lt;0.05,** p&lt;0.01, *** p&lt;0.001. (F) QRT-PCR of CDKN1Bin HCs when CDKN1B was overexpressed. (n=9,3 donors for three replicates) ** p&lt;0.01.(G) MRNA levels of Mmp3, Mmp13, Sox9, Aggrecan, p16INK4a, p21 and p53assessed by qRT-PCR when AC006064.4-201 was overexpressed or co-overexpressedwith CDKN1B. (n=9, 3 donors for threereplicates) * p&lt;0.05, ** p&lt;0.01, *** p&lt;0.001. Supplementary Figure 3. (A) Sequence ofAC006064.4-201. (Red sequence, same part as Gm49317-201 sequence) (B) Knock down efficiency of Gm49317-201 ASOs assessed by qRT-PCR. (n=9, 3 donors for three replicates) ** p&lt;0.01. (C) Western blot ofPTPB1 in MCs aftering pulled-down with the biotinylated Gm49317-201 mixed probes. (D) Knock down efficiency of CDKN1B shRNAs assessed by qRT-PCR. (n=9, 3 donors for three replicates) *** p&lt;0.001. Supplementary Figure 4. (A) Representative images and flurence intensities of IF staining forCDKN1B and Col2a1 in mice cartilage of different groups. (n=10) * p&lt;0.05, ** p&lt;0.01, *** p&lt;0.001. Scale bars, 200µm and 50µm. Supplementary Figure 5. (A) Representative images and flurence intensities of IF staining forCDKN1B and Col2a1 in mice cartilage of different groups. (n=10) * p&lt;0.05, ** p&lt;0.01. Scale bars, 200µm and 50µm.