Journal: Frontiers in Medicine
Article Title: Chaperonin-Containing TCP1 Subunit 5 Protects Against the Effect of Mer Receptor Tyrosine Kinase Knockdown in Retinal Pigment Epithelial Cells by Interacting With Filamentous Actin and Activating the LIM-Kinase 1/Cofilin Pathway
doi: 10.3389/fmed.2022.861371
Figure Lengend Snippet: Upregulation of CCT5 expression recovered the morphology and migration function disrupted by MERTK siRNA in HsRPE cells. (A) HsRPE cells were treated with NC, CCT5 siRNA1, or CCT5 siRNA2 (20 nM) for 24 h, and CCT5 expression was analyzed by RT-PCR. (B) HsRPE cells were treated with or without CCT5 siRNA or CCT5 -specific lentiviral vectors ( CCT5 -Le) after exposure to NC or MERTK siRNA for 24 h, and the expression of CCT5 was analyzed by RT-PCR. (C) HsRPE cells were treated with or without CCT5 siRNA or CCT5 -specific lentiviral vectors ( CCT5 -Le) after exposure to NC or MERTK siRNA for 48 h, and CCT5 expression was analyzed by Western blotting. (D) HsRPE cells were treated the same as in (C), and cell proliferation was determined by CCK8 assay. (E) HsRPE cells were treated the same as in (C) , and morphological changes were observed using an inverted microscope. Scale bars: 100 μm. (F) HsRPE cells were treated the same as in (C) , and wound-healing assays were performed to assess the wound-healing capabilities of HsRPE cells. Scale bars: 200 μm. (G) HsRPE cells were treated the same as in (C) , and Transwell assays were performed to evaluate the migration activity of HsRPE cells (purple). Scale bars: 100 μm. # P > 0.05; * P < 0.05; ** P < 0.01; *** P < 0.001.
Article Snippet: The siRNAs for MERTK and CCT5 were designed and synthesized by RiboBio Inc. (Guangzhou, China).
Techniques: Expressing, Migration, Reverse Transcription Polymerase Chain Reaction, Western Blot, CCK-8 Assay, Inverted Microscopy, Activity Assay
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