- Tubulin Folding Cofactor TBCB is a Target of the Salmonella Effector Protein SseK1

TBCB interacts with SseK1 in bacteria and host cells. (A) Bacterial lysates from strain SV7071 (14028 sseK1::3xFLAG) expressing glutathione S-transferase (GST) or GST-TBCB were prepared in NP40 buffer. Then, purified GST fusion proteins were blotted on nitrocellulose membranes, stained with Ponceau S red (left panel), and developed with monoclonal anti-FLAG (right panel). Aliquots of the original lysates were also included. The molecular weights of the marker bands (M) are indicated. (B) HeLa cells were transiently transfected with a derivative of pBABEpuro expressing SseK1-3xFLAG, a derivative of pCS2 expressing 3xHA-TBCB or cotransfected with the same plasmid and a derivative of pBABEpuro expressing SseK1-3xFLAG. NP40 lysates from 6 × 10⁶ transfected cells were subjected to immunoprecipitation (IP) with anti-FLAG monoclonal antibodies and, after washing, resolved by SDS-PAGE, transferred to nitrocellulose membranes, and developed with monoclonal anti-HA. Total lysates from 5 × 10⁵ transfected cells are included in the blot. M, molecular weight marker. Molecular weights in kDa are indicated on the left.

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Tubulin Folding Cofactor TBCB is a Target of the Salmonella Effector Protein SseK1 - Scientific Figure on ResearchGate. Available from: https://www.researchgate.net/figure/TBCB-interacts-with-SseK1-in-bacteria-and-host-cells-A-Bacterial-lysates-from-strain_fig2_341137384 [accessed 23 Jun 2025]

Caption

TBCB interacts with SseK1 in bacteria and host cells. (A) Bacterial lysates from strain SV7071 (14028 sseK1::3xFLAG) expressing glutathione S-transferase (GST) or GST-TBCB were prepared in NP40 buffer. Then, purified GST fusion proteins were blotted on nitrocellulose membranes, stained with Ponceau S red (left panel), and developed with monoclonal anti-FLAG (right panel). Aliquots of the original lysates were also included. The molecular weights of the marker bands (M) are indicated. (B) HeLa cells were transiently transfected with a derivative of pBABEpuro expressing SseK1-3xFLAG, a derivative of pCS2 expressing 3xHA-TBCB or cotransfected with the same plasmid and a derivative of pBABEpuro expressing SseK1-3xFLAG. NP40 lysates from 6 × 10⁶ transfected cells were subjected to immunoprecipitation (IP) with anti-FLAG monoclonal antibodies and, after washing, resolved by SDS-PAGE, transferred to nitrocellulose membranes, and developed with monoclonal anti-HA. Total lysates from 5 × 10⁵ transfected cells are included in the blot. M, molecular weight marker. Molecular weights in kDa are indicated on the left.

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<a href="https://www.researchgate.net/figure/TBCB-interacts-with-SseK1-in-bacteria-and-host-cells-A-Bacterial-lysates-from-strain_fig2_341137384"><img src="https://www.researchgate.net/publication/341137384/figure/fig2/AS:11431281386787145@1745093127326/TBCB-interacts-with-SseK1-in-bacteria-and-host-cells-A-Bacterial-lysates-from-strain.tif" alt="TBCB interacts with SseK1 in bacteria and host cells. (A) Bacterial lysates from strain SV7071 (14028 sseK1::3xFLAG) expressing glutathione S-transferase (GST) or GST-TBCB were prepared in NP40 buffer. Then, purified GST fusion proteins were blotted on nitrocellulose membranes, stained with Ponceau S red (left panel), and developed with monoclonal anti-FLAG (right panel). Aliquots of the original lysates were also included. The molecular weights of the marker bands (M) are indicated. (B) HeLa cells were transiently transfected with a derivative of pBABEpuro expressing SseK1-3xFLAG, a derivative of pCS2 expressing 3xHA-TBCB or cotransfected with the same plasmid and a derivative of pBABEpuro expressing SseK1-3xFLAG. NP40 lysates from 6 × 10⁶ transfected cells were subjected to immunoprecipitation (IP) with anti-FLAG monoclonal antibodies and, after washing, resolved by SDS-PAGE, transferred to nitrocellulose membranes, and developed with monoclonal anti-HA. Total lysates from 5 × 10⁵ transfected cells are included in the blot. M, molecular weight marker. Molecular weights in kDa are indicated on the left."/></a>