rabbit anti rab8a  (Proteintech)

Journal: bioRxiv

Article Title: The tubulin poly-glutamylase complex, TPGC, is required for phosphatidyl inositol homeostasis and cilium assembly and maintenance

doi: 10.1101/2025.03.03.641315

Figure Lengend Snippet: A ) sgRNA control and TBC1D19 knockout cells were serum starved for 24 hr and immuno-stained with GT335, anti-acetylated tubulin and anti-Arl13b antibodies. ≥90 cells per sample were analyzed in three independent experiments. Error bars, S.D. ***p ≤ 0.001, **** p ≤ 0.0001. B ) sgRNA control and TBC1D19 knockout cells were serum starved for 24 hr, and immuno-stained with antibodies against GT335, Arl13b, Myo-Va, Cep89, Rab8a, Rabin8, IFT88, Cep290 and RPGRIP1L. Scale bar, 1µm. C ) sgRNA control and TBC1D19 knockout cells were serum starved as in panels A and B and visualized by staining with GT335 and antibodies against CP110. Scale bar, 1µm. N ≥ 90 cells per sample were analyzed in three independent experiments. Error bars, S.D. D ) sgRNA control and TBC1D19 knockout cells were serum starved for 24 hr, and immunostained with antibodies against Rab34 and Arl13b. Scale bar, 1µm. E ) sgRNA control and TBC1D19 knockout cells were serum starved as in panels A-C and immunostained with antibodies against GT335 and Arl13b. Scale bar, 1µm. F ) Primary cilia in sgRNA control and TBC1D19 knockout cells were examined by transmission electron microscopy (TEM) after 24 hr of serum starvation. Images of TBC1D19 knockout cilia from two consecutive sections are shown.

Article Snippet: Mouse anti-α-tubulin (1:10000 for WB 66031-1-Ig), rabbit anti-Arl13b (1:2000 for IF and 1:1000 for WB, 17711-1-AP), rabbit anti-Rab8a (1:50 for IF, 55296-1-AP), rabbit anti-IFT88 (1:500 for IF, 13967-1-AP), rabbit anti-INPP5E (1:1000 for WB and 1:1000 for IF, 17797-1-AP), mouse anti-β-actin (1:10000 for WB, 66009-1-Ig PT), mouse anti-GAPDH (1:10000 for WB, 60004-1-Ig), rabbit ant-CCP1 (1:1000 for WB, 14067-1-AP), Rabbit anti-TBC1D19 (1:1000 for WB, 21085-1-AP) and rabbit anti-Tulp3 (1:1000 for WB, 1:150 for IF, 13637-1-AP), RPGRIP1L (1:150 for IF, 55160-1-AP) were from Proteintech.

Techniques: Control, Knock-Out, Staining, Transmission Assay, Electron Microscopy

Journal: bioRxiv

Article Title: The tubulin poly-glutamylase complex, TPGC, is required for phosphatidyl inositol homeostasis and cilium assembly and maintenance

doi: 10.1101/2025.03.03.641315

Figure Lengend Snippet: A ) Control (sgCTL) and TBC1D19 knockout cells were treated with non-targeting control or CCP1 siRNA for 48 hr and serum starved for 24 hr, and extracts were subjected to immunoblotting with the indicated antibodies. B ) TBC1D19 knockout cells were infected with lentivirus expressing Flag-map4m or Flag-map4m-TTLL1 for 72 hr, serum starved for 24 hr and subjected to immunoblotting with the indicated antibodies. C ) Control (sgCTL), TBC1D19 knockout cells, and TBC1D19 knockout cells stably expressing TTLL5-EYFP or TTLL6-EYFP were serum starved for 24 hr and subjected to immunoblotting with the indicated antibodies. D ) Control (sgCTL), TBC1D19 -/- , C11ORF49 -/- , and TTLL1 -/- cells were serum starved for 24 hr and subjected to immunoblotting with the indicated antibodies. E ) sgCTL, C11ORF49 -/- , and LRRC49 -/- cells were serum starved for 24 hr, immuno-stained with antibodies against acetylated tubulin and INPP5E. N ≥ 50 cilia were counted per sample in two independent experiments. Error bars, S.D. ns, not significant. F ) 293T cells were co-transfected with GFP or GFP-TBC1D19 with mCherry-Arl13b for 32 hr and serum starved for 16 hr. Lysates were subjected to immunoprecipitation with GFP-trap beads and immuno-blotted with antibodies against GFP and mCherry.

Article Snippet: Mouse anti-α-tubulin (1:10000 for WB 66031-1-Ig), rabbit anti-Arl13b (1:2000 for IF and 1:1000 for WB, 17711-1-AP), rabbit anti-Rab8a (1:50 for IF, 55296-1-AP), rabbit anti-IFT88 (1:500 for IF, 13967-1-AP), rabbit anti-INPP5E (1:1000 for WB and 1:1000 for IF, 17797-1-AP), mouse anti-β-actin (1:10000 for WB, 66009-1-Ig PT), mouse anti-GAPDH (1:10000 for WB, 60004-1-Ig), rabbit ant-CCP1 (1:1000 for WB, 14067-1-AP), Rabbit anti-TBC1D19 (1:1000 for WB, 21085-1-AP) and rabbit anti-Tulp3 (1:1000 for WB, 1:150 for IF, 13637-1-AP), RPGRIP1L (1:150 for IF, 55160-1-AP) were from Proteintech.

Techniques: Control, Knock-Out, Western Blot, Infection, Expressing, Stable Transfection, Staining, Transfection, Immunoprecipitation