Journal: Experimental & Molecular Medicine

Article Title: Keratin 18 functions as a lactyltransferase to trigger necroptosis in diabetic kidney disease by modulating Fas transcription

doi: 10.1038/s12276-026-01737-9

Figure Lengend Snippet: a Representative microscopy images showing hematoxylin and eosin (HE) and Masson’s trichrome staining of kidney biopsy samples from control (Con) and diabetic kidney disease (DKD) model mice (scale bar, 50 μm). HE staining of renal tissue from DKD model mice revealed marked glomerular enlargement and adaptive tubular hypertrophy. Moreover, Masson’s trichrome staining revealed greater fibrosis in the renal tissues of DKD model mice than in those of control mice (data are presented as the mean ± SD; n = 8 per group). b The levels of kidney metabolites in DKD model mice were normalized to those in control mice and are shown in heatmaps, with the color scale indicating relative differences ( n = 5 in the Con group and n = 4 in the DKD group). c Lactate levels were increased in DKD model mice compared with control mice (data are presented as the mean ± SD; n = 8 per group). d Quantitative PCR assays indicated that the mRNA levels of lactate dehydrogenase A (LDHA) were increased in the kidneys of DKD model mice (data are presented as the mean ± SD; n = 8 per group). e Western blotting revealed that the protein levels of H3K18la, H3K27la, and LDHA were increased in the kidneys of DKD model mice; however, H3K9la levels did not change significantly. f Representative images of immunohistochemistry (IHC) staining for LDHA, Kla, H3K18la, and H3K27la in renal biopsy samples from control group and DKD model mice (scale bar, 50 μm). The levels of LDHA, Kla, H3K18la, and H3K27la were increased in the kidneys of DKD model mice (data are presented as the mean ± SD; n = 8 per group). g Western blotting revealed that the protein levels of Kla were increased in the kidneys of DKD model mice. h Double immunofluorescence staining for Kla and histones H1 (H1), H2A, H2B, H3, or H4 verified the strong colocalization of Kla with H3 in the kidneys of DKD model mice, which indicated that histone lactylation modifications predominantly occur on H3 in DKD. i Representative images of HE staining, Masson’s trichrome staining, and IHC staining for LDHA, Kla, H3K18la, and H3K27la in renal biopsy specimens from patients with DKD in the present study (scale bar, 50 μm). HE staining indicated that glomerular sclerosis intensified progressively and that tubulointerstitial lesions transitioned from multifocal to diffuse involvement as the DKD stage progressed. Masson’s trichrome staining revealed an increase in collagen deposition and interstitial fibrosis as the DKD stage progressed. IHC data indicated that LDHA, Kla, H3K18la, and H3K27la levels in renal biopsy samples from patients with DKD gradually increased with increasing DKD stage (data are presented as the mean ± SD; n = 5 per group). DAPI, 4′,6-diamidino-2-phenylindole; ns, not significant. * P < 0.05 and ** P < 0.01.

Article Snippet: The lactate dehydrogenase A (LDHA) inhibitor OXA (HY-W013032A; MCE) was used in the cellular experiments.

Techniques: Microscopy, Staining, Control, Real-time Polymerase Chain Reaction, Western Blot, Immunohistochemistry, Double Immunofluorescence Staining

Journal: Experimental & Molecular Medicine

Article Title: Keratin 18 functions as a lactyltransferase to trigger necroptosis in diabetic kidney disease by modulating Fas transcription

doi: 10.1038/s12276-026-01737-9

Figure Lengend Snippet: a Oxamate (OXA) treatment decreased lactate accumulation in the renal tissues of diabetic kidney disease (DKD) model mice (data are presented as the mean ± SD; n = 8 per group). b OXA treatment decreased lactate dehydrogenase A (LDHA) mRNA levels in the renal tissues of DKD model mice (data are presented as the mean ± SD; n = 8 per group). c OXA treatment decreased LDHA, H3K18la, and H3K27la levels in the renal tissues of DKD model mice. d OXA treatment reduced lysine lactylation (Kla) levels in the renal tissues of DKD model mice. e Representative images of hematoxylin and eosin (HE) staining, Masson’s trichrome staining and immunohistochemistry staining for LDHA, Kla, H3K18la, and H3K27la in the renal tissues of control (Con), oxamate (OXA), DKD, and DKD + OXA model mice (scale bar, 50 μm). OXA treatment attenuated renal injury and fibrosis and decreased LDHA, Kla, H3K18la, and H3K27la levels in the renal tissues of DKD model mice (data are presented as the mean ± SD; n = 8 per group). f OXA treatment decreased lactate accumulation in hyperglycemic HK-2 cells (data are presented as the mean ± SD; n = 5 per group). g OXA treatment decreased LDHA mRNA levels in hyperglycemic HK-2 cells (data are presented as the mean ± SD; n = 5 per group). h OXA treatment decreased LDHA, H3K18la, and H3K27la levels in hyperglycemic HK-2 cells. i OXA treatment reduced Kla levels in hyperglycemic HK-2 cells. j OXA treatment inhibited the death of hyperglycemic HK-2 cells. HG, high glucose. * P < 0.05 and ** P < 0.01.

Article Snippet: The lactate dehydrogenase A (LDHA) inhibitor OXA (HY-W013032A; MCE) was used in the cellular experiments.

Techniques: Staining, Immunohistochemistry, Control

Journal: Nature Communications

Article Title: TBL1X/TBL1XR1 govern β-cell identity through a PAX6-containing gene regulatory network

doi: 10.1038/s41467-026-72077-5

Figure Lengend Snippet: a Scheme for the generation of an inducible β-cell-specific TBL1X and TBL1XR1 knockout. b Random fed blood glucose levels over time in iTBL/RβKO ( n = 9) and control ( n = 9) mice on HFD. c Total pancreas insulin content normalized to protein levels in iTBL/RβKO and control mice 4 and 11 weeks after tamoxifen administration. n = 3 for 4w control and iTBL/RβKO mice, n = 6 for 11w control mice, n = 4 for 11w iTBL/RβKO mice. d , e Blood glucose ( d ) and plasma insulin ( e ) levels during an oral glucose tolerance test after 22 weeks of HFD. Corresponding area under the curve in iTBL/RβKO ( n = 8) and control ( n = 9) mice ( d , right). f , g Relative mRNA expression determined by qPCR in pancreatic islets of iTBL/RβKO and control mice of Tbl1x and Tbl1xr1 ( f ) and islet genes ( g ). Control mice Tbl1x , Tbl1xr1 , Ins1 , Ins2 , Nkx6.1 , Slc2a2 , Mafa , Pdx1 , Pax6 , Ucn3 : n = 9, control mice ChgA , Ngn3 , Ldha , Hk1 : n = 8, iTBL/RβKO mice Tbl1x , Ngn3 , Ldha , Hk1 : n = 7, iTBL/RβKO mice Tbl1xr1 , Ins1 , Ins2 , Slc2a2 , Mafa n = 6, iTBL/RβKO mice Nkx6.1 , Pdx1 , Pax6 , Ucn3 , ChgA : n = 5. h α/β-cell mass ratio of pancreatic islets from iTBL/RβKO ( n = 4) and control mice ( n = 4) on HFD, 24 weeks after knockout induction. i Representative immunofluorescent staining of insulin + (blue, β-cells) and glucagon + (red, α-cells) cells of paraffin-embedded pancreas from iTBL/RβKO and control mice on HFD, 24 weeks after knockout induction. Each point represents one mouse. Data are represented as mean ± SEM. The following statistical tests were applied: two-sided student’s t test ( d – Area under the curve, f – h ) and 2-way ANOVA with Šidák’s multiple comparison post hoc test ( b – d – time course, e ). Source data are provided as a Source Data file.

Article Snippet: The following TaqMan probes were used: Tbp - Mm01277042_m1, Tbl1x - Mm01222202_m1, Tbl1xr1 - Mm01283877_m1, Ins1 - Mm01259683_g1, Ins2 - Mm00731595_gH, Nkx6.1 - Mm00454961_m1, Slc2a2 - Mm00454961_m1, Mafa - Mm00845206_s1, Pdx1 - Mm00435565_m1, Pax6 - Mm00443081_m1, Ucn3 - Mm00453206_s1, ChgA - Mm00514341_m1, Ngn3 - Mm00437606_s1, Ldha - Mm01612132_g1, Hk1 - Mm00439344_m1, TBP - Hs00427620_m1, TBL1X - Hs00959540_m1, TBL1XR1 - Hs00226564_m1.

Techniques: Knock-Out, Control, Clinical Proteomics, Expressing, Staining, Comparison

Journal: JCI Insight

Article Title: Lactate programs CRIP1 protein lactylation to drive synovial proliferation in rheumatoid arthritis

doi: 10.1172/jci.insight.200928

Figure Lengend Snippet: ( A and B ) IHC staining and quantification of LDHA in synovial tissues from patients with RA and healthy controls ( n = 6). Scale bar: 200 μm. ( C ) Immunoblot analysis of LDHA in synovial tissues from patients with RA and healthy controls ( n = 6). ( D – F ) Immunofluorescence staining and quantification of LDHA (green) and FAPα (red) in synovial tissues from patients with RA and healthy controls ( n = 6). Scale bar: 200 μm. ( G and H ) EdU incorporation assay and quantification of RA-FLS treated with FX11 (10 μM, 20 μM) ( n = 6). Scale bar: 100 μm. ( I ) CCK8 assay to assess proliferation cells treated with FX11 (10 μM, 20 μM) ( n = 6). ( J ) Immunoblot analysis of Pan-Kla in synovial tissues upon FX11 treatment ( n = 3). ( K ) Establishment of a CIA mouse model. Mice were immunized s.c. with type II collagen emulsified in complete Freund’s adjuvant (CII + CFA) on day 0 and again on day 21. FX11 (1 mg/kg, Qd) or vehicle was administered i.p. on day 21. Mice were sacrificed on day 36 for treatment evaluation. ( L ) Immunoblot analysis of Pan-Kla in synovial tissues from HC and CIA mice ( n = 3). ( M ) Immunoblot analysis of Pan-Kla in synovial tissues upon FX11 treatment ( n = 3). ( N and O ) Clinical arthritis score and paw thickness upon FX11 treatment ( n = 6). ( P ) Histological staining with H&E and Safranin O in synovial tissues upon FX11 treatment ( n = 6). Scale bar: 100 μm. * P < 0.05, ** P < 0.01, *** P < 0.001, *** P < 0.001. Data are presented as mean ± SEM, and P values are calculated using unpaired 2-tailed t test ( B , E , and F ), or 1-way ANOVA followed by Tukey’s post hoc test ( H ), or 2-way ANOVA with Bonferroni’s post hoc test ( I , N , and O ).

Article Snippet: In contrast, a selective LDHA inhibitor (FX11; MCE, HY-16214) was applied at 10 μM or 20 μM to inhibit lactylation.

Techniques: Immunohistochemistry, Western Blot, Immunofluorescence, Staining, CCK-8 Assay, Adjuvant

Journal: JCI Insight

Article Title: Lactate programs CRIP1 protein lactylation to drive synovial proliferation in rheumatoid arthritis

doi: 10.1172/jci.insight.200928

Figure Lengend Snippet: ( A ) Schematic workflow of lactylation proteomics in synovial tissues from patients with RA and healthy controls. ( B ) Summary of identified and quantified lactylation sites, peptides, and proteins. ( C ) Subcellular distribution of lactylated proteins. ( D ) Radar chart of 25 representative lactylated proteins with elevated modification levels in patients with RA. ( E and F ) Co-IP of CRIP1 followed by immunoblotting for Pan-Kla to detect CRIP1 protein lactylation in human ( E ) and mouse synovium ( F ). ( G ) Co-IP of CRIP1 followed by immunoblotting for Pan-Kla to detect CRIP1 protein lactylation in FLSs from patients with RA and healthy controls ( n = 3). ( H ) Immunofluorescence staining of CRIP1 (red), Pan-Kla (green), PDPN (yellow), FAPα (cyan) in RA synovium. Scale bar: 100 μm. ( I ) Co-IP of CRIP1 followed by immunoblotting for Pan-Kla to detect CRIP1 lactylation in RA-FLSs after Nala treatment (10 mM, 20 mM) ( n = 3). ( J ) Co-IP of CRIP1 followed by immunoblotting for Pan-Kla to detect CRIP1 lactylation in RA-FLSs after LDHA inhibition with FX11 (10 μM, 20 μM) ( n = 3).

Article Snippet: In contrast, a selective LDHA inhibitor (FX11; MCE, HY-16214) was applied at 10 μM or 20 μM to inhibit lactylation.

Techniques: Modification, Co-Immunoprecipitation Assay, Western Blot, Immunofluorescence, Staining, Inhibition

Journal: Molecular Medicine Reports

Article Title: LDHA protects vascular endothelial cells from oxidative stress-induced mitochondrial damage via HIF-1α activation and glycolytic reprogramming

doi: 10.3892/mmr.2026.13851

Figure Lengend Snippet: LDHA overexpression mitigates apoptosis and mitochondrial damage in OGD/R-treated VECs. (A) Reverse transcription-quantitative polymerase chain reaction analysis of LDHA mRNA in VECs transfected with vector or over-LDHA. (B) Representative western blot analysis of LDHA with β-actin as loading control. (C) Semi-quantification of LDHA protein expression relative to β-actin. (D) Transmission electron microscopy images of VECs at ×2,500 magnification showing mitochondrial morphology. Red arrows indicate mitochondria (swollen and with disrupted cristae under OGD/R conditions). Scale bar: 500 nm. (E) Western blot analysis of caspase-3, Cyt-c and Mcl-1 in VECs from the indicated groups; β-actin was used as the loading control. (F) Densitometric semi-quantification of relative protein expression normalized to β-actin for caspase-3, Cyt-c and Mcl-1. (G) Flow cytometric analysis of the effects of LDHA overexpression on apoptosis levels in VECs subjected to H 2 O 2 (OGD/R) treatment. Data are presented as the mean ± SD of three independent experiments. *P<0.05 vs. vector or as indicated. H 2 O 2 , hydrogen peroxide; OGD/R, oxygen-glucose deprivation/reperfusion; VECs, vascular endothelial cells; LDHA, lactate dehydrogenase A; WB, western blotting; PI, propidium iodide; FITC, fluorescein isothiocyanate; Cyt-c, cytochrome c .

Article Snippet: Subsequently, the membranes were incubated overnight at 4°C with primary antibodies against the following proteins: LDHA (1:1,000; cat. no. 2012; Cell Signaling Technology, Inc.), caspase-3 (1:5,000; cat. no. ab32351; Abcam), caspase-9 (1:2,000; cat. no. ab202068; Abcam), caspase-8 (1:1,000; cat. no. ab32397; Abcam), cytochrome c (Cyt-c; 1:5,000; cat. no. ab133504; Abcam), Mcl-1 (1:1,000; cat. no. ab32087; Abcam), HIF-1α (1:1,000; cat. no. ab51608; Abcam), sodium-calcium exchanger 1 (NCX1; 1:1,000; cat. no. ab177952; Abcam), sodium-proton exchanger 1 (NHE1; 1:1,000; cat. no. ab67313; Abcam) and monocarboxylate transporter 4 (MCT4; 1:1,000; cat. no. ab308528; Abcam), with β-actin (1:1,000; cat. no. ab8227; Abcam) used as an internal loading control for whole-cell and cytoplasmic proteins, and Lamin B1 (1:20,000; cat. no. 12987–1-AP; Wuhan Sanying Biotechnology) used as a nuclear loading control.

Techniques: Over Expression, Reverse Transcription, Real-time Polymerase Chain Reaction, Transfection, Plasmid Preparation, Western Blot, Control, Expressing, Transmission Assay, Electron Microscopy

Journal: Molecular Medicine Reports

Article Title: LDHA protects vascular endothelial cells from oxidative stress-induced mitochondrial damage via HIF-1α activation and glycolytic reprogramming

doi: 10.3892/mmr.2026.13851

Figure Lengend Snippet: LDHA protects against oxidative stress-induced mitochondrial damage through glycolytic reprogramming in VECs. (A) Representative WB images of caspase-3, Cyt-c and Mcl-1 protein expression in VECs treated with 0.5 mM H 2 O 2 (OGD/R) and overexpressing LDHA, with or without 2-DG. Semi-quantification of (B) caspase-3, (C) Cyt-c and (D) Mcl-1 protein levels normalized to internal controls. Measurement of (E) ROS, (F) MDA and (G) GSH levels in each group. Data are presented as the mean ± SD from three independent experiments. *P<0.05, **P<0.01, ***P<0.001 and ****P<0.0001 . H 2 O 2 , hydrogen peroxide; OGD/R, oxygen-glucose deprivation/reperfusion; VECs, vascular endothelial cells; LDHA, lactate dehydrogenase A; 2-DG, 2-deoxy-D-glucose; WB, western blotting; ROS, reactive oxygen species; MDA, malondialdehyde; GSH, glutathione; Cyt-c, cytochrome c .

Article Snippet: Subsequently, the membranes were incubated overnight at 4°C with primary antibodies against the following proteins: LDHA (1:1,000; cat. no. 2012; Cell Signaling Technology, Inc.), caspase-3 (1:5,000; cat. no. ab32351; Abcam), caspase-9 (1:2,000; cat. no. ab202068; Abcam), caspase-8 (1:1,000; cat. no. ab32397; Abcam), cytochrome c (Cyt-c; 1:5,000; cat. no. ab133504; Abcam), Mcl-1 (1:1,000; cat. no. ab32087; Abcam), HIF-1α (1:1,000; cat. no. ab51608; Abcam), sodium-calcium exchanger 1 (NCX1; 1:1,000; cat. no. ab177952; Abcam), sodium-proton exchanger 1 (NHE1; 1:1,000; cat. no. ab67313; Abcam) and monocarboxylate transporter 4 (MCT4; 1:1,000; cat. no. ab308528; Abcam), with β-actin (1:1,000; cat. no. ab8227; Abcam) used as an internal loading control for whole-cell and cytoplasmic proteins, and Lamin B1 (1:20,000; cat. no. 12987–1-AP; Wuhan Sanying Biotechnology) used as a nuclear loading control.

Techniques: Expressing, Western Blot