Skip to main content
. 2023 May 28;10(22):2300953. doi: 10.1002/advs.202300953

Figure 8.

Figure 8

In NSCLC, cigarette smoking is associated with activation of the SOCS2/JAK‐STAT signaling axis and increases of circEML4 in M2‐TAMs, plasma EVs, and NSCLC tissues. A) Immunofluorescence staining for CD163 (red) and circEML4 (green) was measured (n = 8), and B) measurement of CD163 and circEML4 double‐positive cells in NSCLC tissues from smoking or never‐smoking patients (n = 8, randomly selected fields per group). DAPI labels nuclei (blue). Scale bar, 50 µm. C) The shape of EVs was identified by TEM of purified plasma EVs from smoking or never‐smoking NSCLC patients. Scale bar, 200 nm. D) Western blots were performed for EVs. E) By qRT‐PCR, levels of circEML4 were assessed in NSCLC plasma EVs harvested from smoking or never‐smoking patients (n = 20). NSCLC patients with a smoking history were divided into groups according to levels of cigarette smoking (Non‐smokers, n = 48; Mild group, n = 23; Medium group, n = 23; Heavy group, n = 24). F) Levels of circEML4 were assessed by qRT‐PCR in NSCLC tissues from smoking or never‐smoking patients. G) Correlation of circEML4 expression in NSCLC tissues with levels of cigarette smoking by NSCLC patients. H) Western blots were conducted, and I) relative protein expression of SOCS2 was measured (n = 6). J) By qRT‐PCR, expression of SOCS2 was measured in NSCLC tissues from smoking or never‐smoking patients. K) Correlation of SOCS2 expression in NSCLC tissues with levels of cigarette smoking for NSCLC patients. L) Correlation analyses of expression between circEML4 and SOCS2 in NSCLC tissues from smoking or never‐smoking patients. M) IHC staining for p‐JAK2, p‐STAT5, and SOCS2 was performed (n = 8), and N) p‐JAK2, p‐STAT5, and SOCS2 positive signals in NSCLC tissues from smoking or never‐smoking patients were measured (n = 8, randomly selected fields per group). Scale bar, 50 µm. O) Schematic illustration of the regulation of smoke‐promoted progression of NSCLC by stimulation of the ALKBH5/SOCS2/JAK‐STAT signaling axis via EVs circEML4 from M2‐TAMs. All data represent mean ± SD. *p < 0.05; ** p < 0.01.