Fig 2: JPT1 knockdown does not alter the response of endometrial cancer cell lines to metformin. RL95-2 (A) and ACI-181 (B) cells were treated with metformin 72 h following siRNA transfection and dose-response was assessed after an additional 72 h by MTS assay. Proliferation (% control) data represent three technical replicates from one biological sample. Error bars represent standard deviation. Two-way ANOVA was performed after log transformation of the data (P-value was .386639 and .979984 for RL95-2 and ACI-181, respectively)

Fig 3: Jupiter microtubule-associated homolog 1 (JPT1) protein is elevated in metformin responders and decreases following metformin treatment. JPT1 was observed as significantly abundant in metformin responder vs nonresponder patients and was significantly decreased in abundance following metformin treatment in responders by LC-MS/MS-based proteomic analyses (P-values calculated using edgeR testing). The horizontal line inside the box represents the median. The limits of the box show the upper and lower quartiles and the whiskers correspond to the minimum and maximum values observed

Fig 4: Verification of altered JPT1 protein abundance by immunohistochemical analyses of EEC patients' tissues collected from metformin responder (n = 13) vs nonresponders (n = 7). IHC H-Score for JPT1 abundance in metformin responders and nonresponder, pre- and post-metformin treatment (P-values calculated using Mann-Whitney U test). The horizontal line inside the box represents the median. The limits of the box show the upper and lower quartiles and the whiskers correspond to the minimum and maximum values observed

Fig 5: Immunoblots of RL95-2 and ACI-181 demonstrating siJPT1 knockdown. JPT1 expression was knocked down in RL95-2 (A and B) and ACI-181 (C and D) cells by small interfering RNA (siRNA) targeting JPT1 or with a nontargeting (siNT) control and confirmed at the protein (A and B) level before (72 h) and following (168 h) dose-response analyses with metformin treatment. Please note that assessment of MYC abundance in RL-95-2 was generated on a different immunoblot than JPT1, pAKT (S473), and total AKT, but identical protein lysates were used for all RL-95-2 immunoblots. Uncropped immunoblots are shown in Figure S4K-T and aa