Fig. 4

The effect of hsa-miR-21-5p knockdown in microglial cells during CHPV infection. The hsa-miR-21-5p was knockdown and CHPV infection was given in human microglial cells. A The knockdown of hsa-miR-21-5p was confirmed by qPCR using TaqMan primers and probes specific for hsa-miR-21-5p. The expression of anti-miR-21-5p was normalized to the expression of RNU6B. The hsa-miR-21-5p decreased by 70 % in anti-miR-21-5p + CHPV compared to CHPV, while anti-miR-21-5p itself was 30 % downregulated compared to Cy3. B Western Blot analysis showed that PTEN expression was restored following knockdown of hsa-miR-21-5p. C The graph displays the densitometry analysis of PTEN normalized to β-tubulin and showed significant recovery of 20 % in anti-miR-21-5p + CHPV compared to CHPV, while anti-miR-21-5p alone increased the PTEN expression by 30 % compared to Cy3. D Western Blot analysis showed restoration of p-AKT/AKT expression post knockdown of hsa-miR-21-5p and CHPV infection in human microglial cells. E The graph of densitometry analysis of p-AKT/AKT showed significant decrease of 50 % in p-AKT expression in anti-miR-21-5p + CHPV compared to CHPV, while p-AKT/AKT expression in anti-miR-21-5p alone was 28 % downregulated compared to Cy3. The experiments were performed in triplicate (n = 3) and shown as SE ± mean. One-way ANOVA has been applied using by using GraphPad-prism 8 (ver 8.3.0) to calculate the p-values, where: *p < 0.05, **p < 0.01, and ***p < 0.001