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. 2024 Oct 28;13(10):5484–5496. doi: 10.21037/tcr-24-248

Figure 3.

Figure 3

ISL1 promotes the stemness of GC cells in vitro. Representative images of exogenous ISL1 overexpressing (A,B) or knockdown ISL1 (C,D) cells cultured in serum-free medium for 10 days (biological repeat 3 times), observed and photographed using a microscope at 4× and 20× respectively. And statistical analysis was performed on the number of spheroids (diameter >50 µm). Real-time qRT-PCR was used to detect the expression of CD44 (E), AQP5 (F), SOX2 (G) and OCT4 (H) in ISL1 knockdown cells and negative control cells. qRT-PCR was used to detect the expression of CD44 (I), AQP5 (J), SOX2 (K) and OCT4 (L) in ISL1 overexpressing cells and adenovirus control cells (biological repeat 3 times). (M,N) The protein expression of ISL1, AQP5, SOX2 and GAPDH in ISL1 knockdown or ISL1 overexpressing cells was assayed using Western blotting. Ad vector, empty vector control of exogenous overexpression of ISL1; Ad ISL1, exogenous overexpression of ISL1; sh control, empty vector control of knockdown of ISL1 expression; sh ISL1, knockdown of ISL1 expression, SOX2, SRY-box transcription factor 2; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; CD44, CD44 molecule; OCT4, POU class 5 homeobox 1; AQP5, aquaporin 5; ISL1, insulin gene enhancer binding protein-1; GC, gastric cancer; qRT-PCR, quantitative reverse transcription-polymerase chain reaction.