Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: PHLDB2 Mediates Cetuximab Resistance via Interacting With EGFR in Latent Metastasis of Colorectal Cancer

doi: 10.1016/j.jcmgh.2021.12.011

Figure Lengend Snippet: Antibodies and Reagents

Article Snippet: p-Src (1:1000) , Cell Signaling Technology , 59548.

Techniques: Transfection

Journal: Human Gene Therapy

Article Title: A Modified Arrestin1 Increases Lactate Production in the Retina and Slows Retinal Degeneration

doi: 10.1089/hum.2021.272

Figure Lengend Snippet: Rate of lactate production in retinas from mice expressing reduced levels of arrestin1. (A) Example of time course of lactate secretion from excised, dark-adapted retinas placed in DMEM from Arr1 +/+ , Arr1 +/− , and Arr1 −/− mice; inset shows anti-arrestin1 immunoblot of retinal extracts from the three Arr1 genotypes, demonstrating that arrestin1 expression in Arr1 +/− is ∼50% of wild-type levels, and is not detectable in Arr1 −/− (extracts of retinas from the indicated genotype were probed with C10C10 anti-arrestin1 antibody). (B) Average rates of lactate secretion derived from retinas from (A) ; mean ± SD ( n = 10 retinas, 5 mice). (C) Average rates of glucose consumption of retinas from Arr1 +/+ and Arr1 +/− mice; mean ± SEM ( n = 4 retinas). (D) Lactate dehydrogenase levels in different Arr1 genotypes are unchanged; extracts from three retinas used in (B) for each Arr1 genotype ( Arr1 +/+ , Arr1 +/− , and Arr1 −/− ) were immunoblotted and probed with anti-LDHA (upper blot) or anti-β-tubulin (lower blot). (E) Quantitation of arrestin1 and enolase1 expression in whole retinal extracts of C57BL/6J mice. An example of enolase1 immunoblotting (Eno1) and arrestin1 immunoblotting (Arr1) are shown comparing three different volumes (in μL) of retinal extract with known quantities of purified enolase1 and arrestin1 (in pmoles). (F) Quantitation of multiple blots as in (E) , showing the average concentration of arrestin1 and enolase1 and molar ratios in each retina (bars show mean ± SEM; n = 3 retinas). (G) Inhibition of enolase1 catalysis by arrestin1 (ArrWT) is competitively reduced by addition of ArrGG; the rate of turnover of 2-PGA was monitored in the presence of 50 nM enolase1 without ArrWT ( green dashed line ) or with 100 nM ArrWT ( red dashed line ). Addition of the indicated concentration of ArrGG reduced the suppression of enolase1 catalysis caused by ArrWT (points show mean ± SEM; n = 3). 2-PGA, 2-phosphoglycerate; DMEM, Dulbecco's modified Eagle's medium; LDHA, lactate dehydrogenase A; SD, standard deviation; SEM, standard error of the mean. Color images are available online.

Article Snippet: Duplicate blots were probed with either anti-LDHA (1:1,000; Cell Signaling Technology #2102) or anti-β-tubulin (1 μg/mL Santa Cruz #SC-5274).

Techniques: Expressing, Western Blot, Derivative Assay, Quantitation Assay, Purification, Concentration Assay, Inhibition, Modification, Standard Deviation