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Location Gallstones can grow in different tracts of the biliary tree and their localization governs the choice of the appropriate therapeutic approach

Immunoblot analysis For detection of Gclc (Santa Cruz Biotechnology sc-390811), Pdha-1 (Abcam ab168379), DRP1 (Abcam ab184247), eIF4E (Abcam ab33766), p-eIF4E (Abcam ab76256), 4E-BP1 (Cell signaling 9452), p-4E-BP1 (Cell signaling 2855), DRP1 (Abcam ab184247), mTOR (Cell Signaling 2972), p-mTOR (Ser2448) (Cell Signaling 2971), p70 S6 (Cell Signaling 9202), p-p70 S6 (Thr421/Ser424) (Cell Signaling 9204), 15 10 6 cells were lysed with lysis buffer (CST 9803S) and protein/phosphatases inhibitors as per manufacturers protocol and blotted as described 44

Blood 84(10):147a, 1994

The PCR reaction was carried out in a final volume of 25 l containing 2X PCR master mix (EmeraldAmp GT PCR Master Mix, Takara Bio Inc., Shiga, Japan), 20 pM of each primer (forward primer: 5'TGAAGGAGAAGGTGTCTGCGGGA3' and reverse primer: 5'AGGACGGTGCGGTGAGAGTG3' synthesized from Sigma-Aldrich Chemical Pvt Limited, (Milwaukee, Wis., USA), 100 ng of the DNA template and nuclease-free water