Fig. 5: RCAN3 inhibits perfusion recovery in experimental PAD. | Communications Medicine

Fig. 5: RCAN3 inhibits perfusion recovery in experimental PAD.

From: Inhibiting anti-angiogenic VEGF165b activates a miR-17-20a-Calcipressin-3 pathway that revascularizes ischemic muscle in peripheral artery disease

Fig. 5

a Western blot analysis of RCAN3 in ischemic muscle treated with negative inhibitor (Neg-Inh, blue bar) or a combination of miR-17 and miR-20a inhibitor (miR-17-20 Inh, pink bar). n = 4. Unpaired t-test. b qPCR of RCAN3 expression in C57BL/6J nonischemic (NI, blue bar) and ischemic (I, pink bar) muscle. n = 4. Unpaired t-test. c Laser Doppler perfusion imaging of microvascular blood flow in ischemic muscle transfected with a control plasmid (Neg Pld, blue box plots, n = 4) or RCAN3-expressing plasmid (RCAN3 Pld, pink box plots, n = 5). Repeated-measures two-way ANOVA with Bonferroni’s post-test. d Immunohistochemical analysis of SMA (green) and CD31 (red) in ischemic gastrocnemius muscle treated with Neg Pld (n = 4, blue bars) or RCAN3 Pld (n = 5, pink bars) at day 14 post-HLI. Scale bars are 50 µm. Unpaired t-test. e Immunohistochemical analysis of CD31 (red) and PCNA (green) in ischemic gastrocnemius muscle treated with Neg Pld (n = 4, blue bars) or RCAN3 Pld (n = 5, pink bars) at day 14 post-HLI. Scale bars are 50 µm. Unpaired t-test. P < 0.05 significant. Data from the biological replicates are presented as mean  ±  standard error.

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