Figure 6

Immunofluorescence of FLAG-SK1a channels or PKD2-FLAG channels in BBS7 - or BBS8 - or BBS9 -depleted cells). (A) FLAG-SK1a channels. (B) PKD2-FLAG channels. FLAG-SK1a- expressing cells were stained with anti-FLAG antibody and as a contrast, anti-folate chemoreceptor (FBP) antibody in a series of control and RNAi conditions. Only the merged images are shown here. Complete immunofluorescence of the folate chemoreceptor and the FLAG-SK1a channel are shown separately in Additional file 9: Figure S5. The FLAG control is a cell microinjected with FLAG-pPXV vector and fed with RNAi empty vector (L4440) bacteria. The FLAG-SK1a control is a cell expressing FLAG-SK1a and fed bacteria with an RNAi empty vector (L4440). BBS7, BBS8 and BBS9 are the cells expressing FLAG-SK1a channel and also BBS7-, BBS8- or BBS9-depleted, respectively. Cells were immunostained with anti-FLAG (red) and anti-FBP (green) antibodies. Differential interference contrast (DIC) images are shown to document that cilia are present. Cells expressing PKD2-FLAG channel (B) were stained with anti-FLAG antibody (red) and anti-Tetrahymena centrin-1 antibody (green) in a series of control and RNAi conditions. Only the merged images are shown here; the staining of FLAG and centrin can be seen separately in Additional file 10: Figure S6. The FLAG control is a cell expressing the FLAG-pPXV vector and fed with RNAi empty vector (L4440) bacteria. The PKD2-FLAG control is a cell expressing the PKD2-FLAG channel and fed with bacteria with RNAi empty vector (L4440). BBS7, BBS8 and BBS9 are the cells expressing the PKD2-FLAG channel and are BBS7-, BBS8- or BBS9- depleted, respectively. Differential interference contrast (DIC) images are shown to document that cilia are present. All images were taken under 60× oil immersion objectives. Scales represent 15 μm. Images are representative of results of three experiments, n = 126 to 156 cells.