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. 2021 Jul 27;11:718731. doi: 10.3389/fonc.2021.718731

Figure 5.

Figure 5

Overexpression of miR-142a impaired cell viability partially through targeting Ciapin1. (A) Follow-up analysis of GFP+ cell proportion in 32D-BA cells infected with pCDH-EF1-miR142a-T2A-copGFP virus. (B) Apoptosis analysis of 32D-BA cells at 24h and 48h after lentivirus infection. (C) Validation of miR-142a-5p and miR-142a-3p expression by qRT-PCR. (D, E) qRT-PCR detection of anti-apoptotic gene Bcl2, Birc2, Ciapin1, and Xiap in 32D-BA cells (D) and miR-142a-overexpressed (OE142a) 32D-BA cells (E). (F) Western blot detection of anti-apoptotic proteins Bcl2, Birc2, Ciapin1, and Xiap in 32D-BA cells infected with lentivirus encoding miR-142a. (G) Identification of miR-142a target gene by luciferase reporter assay. (H) Apoptosis analysis of 32D-BA cells at 24h and 48h after Ciapin1 knockdown. (I) Determination of Ciapin1 expression by qRT-PCR and western blot analysis. (J) Comparative analysis of apoptosis between 32D-BA-puro+OE142a and 32D-BA-Ciapin1+OE142a cells via Annexin V staining. (K) Validation of Ciapin1 expression by qRT-PCR and Western blot assay. (L) Validation of miR-142a-5p and miR-142a-3p expression in 32D-BA-puro and 32D-BA-Ciapin1 cells by qRT-PCR. Results are mean ± SEM, *p < 0.05, **p < 0.01, ***p < 0.001 and N/S (not significant) (B–E, G–L) was assessed by unpaired t-test.