Fig. 5

lnc-Helf promotes HMs M1 polarization and proliferation. A–C Mouse primary HMs transfected with siRNA for 24 h, following treatment with 20 ng/ml IFN-γ for 24 h. qRT–PCR analysis was used to detect the RNA level of lnc-Helf, Ly6c, Tnf-α, Mcp-1, Il-1β, Il-6, Pcna, Cyclin D1, and Cyclin E1 (A); mature supernatant IL-1β level was detected by ELISA (B); the expression and location of TNF-α was assessed by confocal microscopy (C). Scale bar is 20 μm. D HMs were transfected with pcDNA3.1-lnc-Helf or pcDNA3.1 for 48 h; qRT–PCR was used to assess the expression of lnc-Helf, Ly6c, Tnf-α, Mcp-1, Il-1β, Il-6, Pcna, Cyclin D1, and Cyclin E1. E, F HMs were transfected with pcDNA3.1-lnc-Helf or pcDNA3.1 for 48 h, following treatment with 20 ng/ml IFN-γ for 24 h. Mature supernatant IL-1β level was detected by ELISA (E); the expression and location of TNF-α was assessed by confocal microscopy (F). Scale bar is 20 μm. G RAW264.7 cells were transfected with pcDNA3.1-lnc-Helf or pcDNA3.1 for the indicated times, cell proliferation was detected by CCK8. Data are presented as mean ± SEM. ^*/#p < 0.05. *p < 0.05 for si-NC or pcDNA3.1. ^#p < 0.05 for si-NC + IFN-γ or pcDNA3.1 + IFN-γ, one-way ANOVA (A, B, E) and unpaired Student’s t test (D, G)