Figure S3.
LUZP1 is an actin-stabilizing protein. (A and B) ARP2 and actin fluorescence intensity at the centrosome of RPE-1 cells stably expressing GFP-only, GFP-LUZP1, GFP-EPLINα, or GFP-EPLINβ. The bar indicates total integrated intensity. The bar indicates average intensity. ***, P < 0.01 (Student’s two-tailed t test).(C and D) Representative images of RPE-1 cells stably expressing GFP-only, GFP-LUZP1, GFP-EPLINα, or GFP-EPLINβ and stained with antibodies against PCNT (centrosome marker) and ARP2 (C), or with phalloidin (D). (E) Western blot analysis of endogenous and LUZP1 and EPLIN in RPE-1 cells transfected with the indicated siRNAs for 72 h. (F) WT RPE-1 cells were transfected for 72 h with a control siRNA or siRNAs targeting LUZP1 or EPLIN, individually or in combination. Bar graph shows the mean percentage of ciliated cells (n > 200 cells per sample, three independent experiments) in each population. Error bars indicate SD. *, P < 0.05 (Student’s two-tailed t test). (G) IF analysis of RPE-1 GFP-EPLINβ cells transfected for 72 h with control siRNA or siRNAs targeting LUZP. During the last 16 h of transfection, cells were treated with either DMSO (control) or 50 nM cytochalasin D. Cells were stained for γ-tubulin (centrosome and basal body marker) and ARL13B (ciliary marker). DNA was stained with DAPI. (H) Western blot analysis using protein extracts prepared from RPE-1 control cell lines and lines expressing GFP-LUZP1 or GFP-EPLINα or β. The cells were transfected with a control siRNA or siRNAs targeting LUZP1 or EPLIN, as indicated. A GFP antibody was used to detect the GFP-fusions and antibodies against LUZP1 and EPLIN were used to detect the respective endogenous proteins.

LUZP1 is an actin-stabilizing protein. (A and B) ARP2 and actin fluorescence intensity at the centrosome of RPE-1 cells stably expressing GFP-only, GFP-LUZP1, GFP-EPLINα, or GFP-EPLINβ. The bar indicates total integrated intensity. The bar indicates average intensity. ***, P < 0.01 (Student’s two-tailed t test).(C and D) Representative images of RPE-1 cells stably expressing GFP-only, GFP-LUZP1, GFP-EPLINα, or GFP-EPLINβ and stained with antibodies against PCNT (centrosome marker) and ARP2 (C), or with phalloidin (D). (E) Western blot analysis of endogenous and LUZP1 and EPLIN in RPE-1 cells transfected with the indicated siRNAs for 72 h. (F) WT RPE-1 cells were transfected for 72 h with a control siRNA or siRNAs targeting LUZP1 or EPLIN, individually or in combination. Bar graph shows the mean percentage of ciliated cells (n > 200 cells per sample, three independent experiments) in each population. Error bars indicate SD. *, P < 0.05 (Student’s two-tailed t test). (G) IF analysis of RPE-1 GFP-EPLINβ cells transfected for 72 h with control siRNA or siRNAs targeting LUZP. During the last 16 h of transfection, cells were treated with either DMSO (control) or 50 nM cytochalasin D. Cells were stained for γ-tubulin (centrosome and basal body marker) and ARL13B (ciliary marker). DNA was stained with DAPI. (H) Western blot analysis using protein extracts prepared from RPE-1 control cell lines and lines expressing GFP-LUZP1 or GFP-EPLINα or β. The cells were transfected with a control siRNA or siRNAs targeting LUZP1 or EPLIN, as indicated. A GFP antibody was used to detect the GFP-fusions and antibodies against LUZP1 and EPLIN were used to detect the respective endogenous proteins.

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