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. 2022 Nov 17;12(11):e1113. doi: 10.1002/ctm2.1113

FIGURE 4.

FIGURE 4

SRSF11 promotes cell migration and invasion via HSPA12A‐Ex2+ stabilizing N‐cadherin mRNA in vivo. (A) Confirmation of the RNA levels of two HSPA12A transcripts after knockdown of HSPA12A treatments in LoVo and SW480 cell lines. (B) The cell migration and invasion of LoVo and SW480 cells after transfection with si‐HSPA12A and the negative control were detected by Transwell assays (N = 3; Student t′ test; **p < .01, ***p < .001). (C) Representative images of the cell wound healing after transfection with si‐HSPA12A and respective negative control (N = 3; Student t′ test; **p < .01). (D) Transwell assay for investigating the abilities of migration and invasion in LoVo and SW480 cells after co‐transfection with indicated plasmid and siRNA (N = 3; Student t′ test; **p < .01, ***p < .001). (E) Representative images of the cell wound healing of shift after co‐transfection with indicated plasmid and siRNA. The statistical diagrams were displayed below (N = 3; Student t′ test; **p < .01, ***p < .001). (F) WB analyses of EMT markers, E‐cadherin and N‐cadherin in LoVo cells after treatment with SRSF11 overexpression or knockdown of SRSF11 or HSPA12A. (G) qPCR detection of N‐cadherin mRNA after treatment with si‐HSPA12A (N = 3; Student t′ test; **p < .01). (H) RNA stability detection between si‐HSPA12A and si‐NC group by ActD treatment for 4 h (N = 3; SNK test, *p < .05, **p < .01, ***p < .001). (I) Representative immunofluorescence images by confocal microscope indicate N‐cadherin (green), Phalloidin (red) and DAPI (blue) localization in LoVo cells. Phalloidin staining was used to observe the actin cytoskeleton. (J) RIP experiments of N‐cadherin RNA by AGO2 antibody in LoVo and SW480 cell lines (N = 3; Student t′ test; *p < .05). (K) RIP experiments of HSPA12A RNA by AGO2 antibody in LoVo and SW480 cell lines (N = 3; Student t′ test; *p < .05, ***p < .001). (L) RIP experiments of N‐cadherin RNA by AGO2 antibody after treatment with si‐NC or si‐HSPA12A in LoVo and SW480 cell lines (N = 3; Student t′ test; *p < .05)