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. 2026 Feb 3;16:7079. doi: 10.1038/s41598-026-38087-5

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© The Author(s) 2026

Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if you modified the licensed material. You do not have permission under this licence to share adapted material derived from this article or parts of it. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by-nc-nd/4.0/.

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Fig. 2 — Obinutuzumab inhibits TRPML2-Mediated Lysosomal Ca²⁺ Release. (a,b) To measure time dependent TRPML2 inhibition, ML-SA1 was treated at indicated time point after 10 μg/ml OBI treatment. Average cytosolic Ca²⁺ increases were observed in control, RTX (10 μg/ml), and OBI (10 μg/ml) treated Ramos cells upon ML-SA1 treatment using Fluo-4 Ca²⁺ dye. (c) Time-course traces of lysosomal Ca²⁺-dependent fluorescence in Ramos cells loaded with Oregon Green BAPTA–dextran. After pretreatment with mock (vehicle) or OBI (10 μg/ml), cells were stimulated with ML-SA1 during the time window indicated. (d) Quantification of the mean Oregon Green BAPTA–dextran fluorescence averaged over the ML-SA1 stimulation window indicated in (c), comparing mock- vs OBI-pretreated cells. (e) Quantification of the mean Oregon Green BAPTA–dextran fluorescence averaged over the antibody-treatment window (indicated in the trace), comparing mock (vehicle) and OBI conditions. Data are presented as mean ± SEM. *p < 0.05, ***p < 0.001 vs the mock (vehicle) control. (e) Average fluorescence changes of Oregon Green BAPTA-dextran under mock conditions and during OBI treatment. (f,i) Evaluation of OBI’s effect on TRPML2 activity in Ramos cells using fluorescence changes of TRPML2-GCaMP6s (f,g) and GCaMP6s-TRPML2^Va (h,i). Following antibody treatment, fluorescence trends of GCaMP were tracked upon administration of 20 μM ML-SA1 at indicated points. Fluorescence changes in GCaMP6s-TRPML2 and GCaMP6s-TRPML2^Va expressing cells were assessed upon ML-SA1 induction compared to control, RTX, and OBI treatment. (j,k) The traces of GCaMP6s-TRPML2 fluorescence showing OBI-time (10 μg/ml) (j) and -dose (k) dependent, achieving full inhibition at 3 μg within 5 min. One-way ANOVA with Dunnett’s multiple-comparisons test (vs. control) used in (b,g,i). Unpaired two-tailed Student’s t-test used in (d,e). Data are presented as mean ± SEM. *p < 0.05, ***p < 0.001 compared to control group. Data are presented as mean ± SEM. *p < 0.05, ***p < 0.001 compared to control group.