Journal: Molecular Medicine Reports

Article Title: Role of IFT88 in icariin-regulated maintenance of the chondrocyte phenotype

doi: 10.3892/mmr.2018.8486

Figure Lengend Snippet: Primer sequences.

Article Snippet: Using standard immunofluorescence methods, the primary cilia were stained with anti-acetylated α-tubulin antibody (cat. no. T7451; 1:300 dilution; Sigma-Aldrich; Merck KGaA, Darmstadt, Germany) or double-stained with that antibody together with an antibody against IFT88 (cat. no. AP11138b; 1:50 dilution; Abgent Inc., San Diego, CA, USA).

Techniques:

Journal: Molecular Medicine Reports

Article Title: Role of IFT88 in icariin-regulated maintenance of the chondrocyte phenotype

doi: 10.3892/mmr.2018.8486

Figure Lengend Snippet: Icariin promotes ciliary assembly and IFT88 expression. (A) Primary cilia were stained for acetylated-α-tubulin (red coloration, denoted by the white arrows; magnification, ×200). (B) The histograms reveal that, compared with the control group, 10 µmol/l icariin increased primary ciliary assembly (Control group, 27.91±9.95% cf. Icariin, 34.06%±10.06; *P<0.05), and icariin moderately increased the ciliary length from 3.07±0.74 to 3.34±1.34 µm. (C) Icariin increased IFT88 gene expression, peaking at 10 µmol/l (*P<0.05 cf. 0 µmol/l icariin). (D) Icariin upregulated production of the ciliary protein, IFT88, in a concentration- and time-dependent manner (*P<0.05 cf. 0 µmol/l icariin, or treatment at 0 h). IFT88, intraflagellar transport protein 88.

Article Snippet: Using standard immunofluorescence methods, the primary cilia were stained with anti-acetylated α-tubulin antibody (cat. no. T7451; 1:300 dilution; Sigma-Aldrich; Merck KGaA, Darmstadt, Germany) or double-stained with that antibody together with an antibody against IFT88 (cat. no. AP11138b; 1:50 dilution; Abgent Inc., San Diego, CA, USA).

Techniques: Expressing, Staining, Concentration Assay

Journal: Molecular Medicine Reports

Article Title: Role of IFT88 in icariin-regulated maintenance of the chondrocyte phenotype

doi: 10.3892/mmr.2018.8486

Figure Lengend Snippet: Icariin enhances primary ciliary assembly and maintains the phenotype of the primary chondrocytes. (A) Primary cilia were stained for acetylated-α-tubulin (red). Functional IFT88 (green) was widely dispersed in the cytoplasm, although it was principally centralized along the ciliary axonemes, which also contained acetylated α-tubulin (denoted by the white arrows; magnification, ×200). (B) Icariin increased the proportion of primary chondrocytes with cilia, from 29.54±8.24 to 38.48±10.36%, and led to a slight elongation of the cilia from 2.48±0.85 to 2.65±0.63 µm (*P<0.05). (C) Toluidine blue staining revealed that icariin promoted cartilage matrix secretion by primary chondrocytes. (D) Expression of phenotype-associated genes in primary chondrocytes (*P<0.05). IFT88, intraflagellar transport protein 88.

Article Snippet: Using standard immunofluorescence methods, the primary cilia were stained with anti-acetylated α-tubulin antibody (cat. no. T7451; 1:300 dilution; Sigma-Aldrich; Merck KGaA, Darmstadt, Germany) or double-stained with that antibody together with an antibody against IFT88 (cat. no. AP11138b; 1:50 dilution; Abgent Inc., San Diego, CA, USA).

Techniques: Staining, Functional Assay, Expressing

Journal: Molecular Medicine Reports

Article Title: Role of IFT88 in icariin-regulated maintenance of the chondrocyte phenotype

doi: 10.3892/mmr.2018.8486

Figure Lengend Snippet: Icariin regulates IFT88 expression via the ERK signalling pathway. (A) Quantitative polymerase chain reaction measuring the expression levels of the IFT88 and various phenotype-associated genes after transformation of IFT88 siRNA (*P<0.05 compared with the control group; ## P<0.01 compared with the icariin group). (B) IFT88 knockdown reduced the expression of phosphorylated ERK (**P<0.01 compared with the siCon group). (C) Icariin promoted expression of IFT88 and phosphorylated ERK, and slightly increased ERK phosphorylation, after transformation of IFT88 siRNA (**P<0.01 compared with the control group). (D) An ERK inhibitor reduced IFT88 expression. Icariin upregulated IFT88 expression, but could not restore such expression in the presence of the ERK inhibitor, PD0325901 (*P<0.05; **P<0.01 compared with the control group). ERK, extracellular signal-regulated kinase; IFT88, intraflagellar transport protein 88; Ica, icariin; PD, PD0325901, Con, control; T-ERK, total ERK; P-ERK, phosphorylated ERK.

Article Snippet: Using standard immunofluorescence methods, the primary cilia were stained with anti-acetylated α-tubulin antibody (cat. no. T7451; 1:300 dilution; Sigma-Aldrich; Merck KGaA, Darmstadt, Germany) or double-stained with that antibody together with an antibody against IFT88 (cat. no. AP11138b; 1:50 dilution; Abgent Inc., San Diego, CA, USA).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Transformation Assay