Fig 1: MiR-136-5p directly targets RAB9A. (a) Venn diagram showing seven possible target genes for miR-136-5p in four databases (Starbase, Targetscan, miRDB, and TarBase v.8). (b and c) The expression of seven possible target genes was detected in A875 and SK-MEL-1 cells transfected with miR-NC or miR-136-5p. (d) The putative binding sequence between miR-136-5p and RAB9A 3′-UTR was displayed. (e and f) Dual-luciferase reporter assay was used to verify the relationship between miR-136-5p and RAB9A. (g and h) The mRNA and protein levels of RAB9A were tested in melanoma tissues and normal tissues. (i and j) The expression of RAB9A in HEMn-LP, A875, and SK-MEL-1 cells was measured by qRT-PCR and western blot. (k) RAB9A protein level was determined in A875 and SK-MEL-1 cells transfected with miR-NC or miR-136-5p. (l and m) The mRNA and protein levels of RAB9A were examined in A875 and SK-MEL-1 cells introduced with si-NC, si-circ_0013359, si-circ_0013359 + anti-miR-NC, or si-circ_0013359 + anti-miR-136-5p. *P < 0.05.
Fig 2: RAB9A overexpression mitigates the inhibitory effect of miR-136-5p on melanoma cell progression. A875 and SK-MEL-1 cells were transfected with miR-NC, miR-136-5p, miR-136-5p + vector, or miR-136-5p + RAB9A, respectively. RAB9A protein level (a), cell viability (b and c), colony numbers (d), apoptosis rate (e), cell cycle progress (f and g), cell migration and invasion (h and i), glucose consumption (j), lactate production (k), ECAR (l and m), and the protein levels of HK2 and LDHA (n and o) were examined via the appropriate methods. *P < 0.05.
Fig 3: Silencing of circ_0013359 inhibits the growth of melanoma xenografts. A875 cells (1 × 106) stably transfected with sh-NC or sh-circ_0013359 were subcutaneously injected into the nude mice. (a) Tumor volume was measured every 7 days. (b) After the mice were killed, the xenograft tumors were weighed. (c) The photos of tumor on day 35 were shown. (d–f) The levels of circ_0013359, miR-136-5p, and RAB9A were measured by qRT-PCR or western blot. *P < 0.05.
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