Figure 3: miR-15 family expression is downregulated by Salmonella infection. | Nature Communications

Figure 3: miR-15 family expression is downregulated by Salmonella infection.

From: Functional high-throughput screening identifies the miR-15 microRNA family as cellular restriction factors for Salmonella infection

Figure 3

(a) Changes in miRNA expression upon Salmonella infection (MOI 25) of HeLa cells. Results are shown for total population, fraction of cells with internalized Salmonella (Salmonella+) and bystander fraction (Salmonella−). Only miRNAs with number of reads ≥25 in the mock-treated sample are shown. The inset shows expression of the miR-15 family members. The colour scale illustrates the frequency of miRNA expression (log2 scale) for the different cell populations (total, Salm− and Salm+). (b,c) Expression of mature miR-15a and miR-16 determined by qRT–PCR in HeLa cells infected with (b) Salmonella wild type at the indicated MOIs or (c) mutant strains defective in invasion (ΔSPI-1, MOI 100), intracellular replication (ΔSPI-2, MOI 100) or heat-killed Salmonella (MOI 100). (d) Quantification of miR-15 pri-miRNA transcripts using qRT–PCR in HeLa cells infected with Salmonella at the indicated MOIs. miR-15a/miR-16-1 and miR-15b/miR-16-2 are processed from DLEU2 and SMC4 transcripts, respectively. (e,f) E2F1 expression determined using western blot upon infection with (e) Salmonella wild type at the indicated MOIs or (f) mutant strains defective in invasion (ΔSPI-1, MOI 100) or intracellular replication (ΔSPI-2, MOI 100). (g,h) Expression of (g) pri-miRNAs and (h) mature forms of miR-15 family members upon knockdown of E2F1, determined using qRT–PCR in HeLa cells. Samples were collected at 20 h.p.i. Results are shown as mean±s.e.m., normalized to mock-treated cells (b–d) or control siRNA (g,h). ***P<0.001, two-tailed Student’s t-test; n≥3.

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